This study established the mouse podocyte clone-5 (MPC5) with Smad3 knockout and studied the effect of transforming growth factor-beta 1 (TGF-β1) on the dedifferentiation of the MPC5 cells with Smad3 knockout, aiming to provide a cell tool for studying the role of Smad3 in mouse podocytes. The single-guide RNA (sgRNA) sequence targeting Smad3 was designed according to the principles of CRISPR/Cas9 design. The pX458-Smad3 vector was constructed and introduced into competent cells, and then the vector was extracted and used to transfect MPC5 cells. The successfully transfected cells were sorted by a flow cytometer. After single-cell clone expansion, PCR amplification of sequences adjacent to the edition site of Smad3 and sequencing were performed to identify potential cells with gene knockout. Western blotting was employed to verify the knockout efficiency of Smad3. Finally, the effect of Smad3 knockout on TGF-β1-induced dedifferentiation of MPC5 cells was analyzed by reverse transcription-polymerase chain reacting (RT-PCR), Western blotting, and the immunofluorescence method. The sgRNA was designed to target the fifth exon of Smad3. EGFP expression was observed 24 h after transfection of the pX458-Smad3 plasmid into MPC5 cells, with the transfection efficiency of 0.1% as determined by flow cytometry. From the transfected cells, 21 cell clones were obtained through flow cytometric sorting and single-cell clone expansion. PCR amplification and sequencing of the region around the sgRNA target site in Smad3 identified two cell clones with biallelic frameshift mutations. Western blotting results confirmed the absence of Smad3 expression in these clones, indicating successful establishment of the MPC5 cell line with Smad3 knockout. In normal MPC5 cells, TGF-β1 stimulation promoted the expression of fibrosis-related genes fibronectin and Col1a1 (collagen I) and inhibited the expression of the podocyte marker proteins synaptopodin and podocin, which suggested epithelial-mesenchymal transition and podocyte injury. However, in the two MPC5 cell lines with Smad3 knockout, TGF-β1-induced expression of epithelial-mesenchymal transition markers was significantly suppressed. The MPC5 cell lines with Smad3 knockout that were constructed by CRISPR/Cas9 provide a valuable cell model for functional studies of Smad3 protein and highlight the critical role of Smad3 in cell dedifferentiation.
Animals ; Smad3 Protein/genetics* ; CRISPR-Cas Systems/genetics* ; Mice ; Podocytes/metabolism* ; Transforming Growth Factor beta1/pharmacology* ; Cell Line ; Gene Knockout Techniques ; RNA, Guide, CRISPR-Cas Systems/genetics*

Objective:To evaluate the effect of preemptive analgesia with ibuprofen on postoperative pain following single posterior tooth implantation, aiming to provide a clinical reference for its application.Methods:A multicenter, randomized, double-blind, placebo-controlled parallel-group trial was conducted. A total of 82 participants were included in the trial, meeting the eligibility criteria from April 2022 to April 2024 at the Capital Medical University School of Stomatology (40 cases), Beijing TianTan Hospital, Capital Medical University (22 cases), Beijing Chao-Yang Hospital, Capital Medical University (20 cases). Participants were randomly assigned in a 1∶1 ratio to either the ibuprofen group or the control group, with each group comprising 41 individuals. Participants in the ibuprofen group received 300 mg of sustained-release ibuprofen capsules orally 15 min before surgery, while the control group received a placebo. Both groups received the same postoperative analgesic regimen for 3 days. Pain scores were assessed using the numerical rating scale at 30 min, 4 h, 6 h, 8 h, 24 h, 48 h, and 72 h postoperatively, and the additional use of analgesic medication was recorded from days 4 to 6 postoperatively.Results:A total of 82 participants were initially enrolled in the study, with 7 dropouts (4 from the control group and 3 from the ibuprofen group), resulting in 75 participants (37 in the control group and 38 in the ibuprofen group) completing the trial. There were no reports of adverse events such as nausea or vomiting among the participants. The ibuprofen group exhibited significantly lower pain scores at 4 h, 6 h and 8 h [1.0 (0.0, 2.0), 1.0 (0.0, 2.0), 1.5 (0.0, 3.0) ] postoperatively compared to the control group 4 h, 6 h and 8 h [2.0 (1.0, 3.0), 3.0 (1.5, 4.0), 2.0 (1.0, 4.0)] ( Z=-1.99, P=0.047; Z=-3.01, P=0.003; Z=-2.10, P=0.036). The proportions of patients requiring additional analgesic medication between days 4 and 6 post-surgery were 18.4% (7/38) in the ibuprofen group and 27.0% (10/37) in the control group, with no significant difference (χ 2=0.79, P=0.373). The median additional medication usage postoperatively was [0.0 (0.0, 0.0) pills] in the ibuprofen group and [0.0 (0.0, 1.0) pills] in the control group, with no significant difference ( Z=-0.78, P=0.439). Conclusions:Preemptive analgesia with ibuprofen effectively reduces postoperative pain following tooth implantation, representing a safe and effective perioperative pain management strategy.

Objective To explore the relationship between preoperative coronary angiography and postoperative acute kidney injury (AKI) in cardiac surgery. Methods The clinical data of patients who underwent coronary angiography within 30 days before cardiac surgery in the First Affiliated Hospital of Xi’an Jiaotong University from January 2015 to April 2019 were retrospectively analyzed. Univariate analysis and multivariate logistic regression analyses were used to explore the relationship between the interval from preoperative coronary angiography to cardiac surgery and postoperative AKI. Results Finally 1 112 patients were collected, including 700 males and 412 females, with a median age of 61 (55, 66) years. The incidence of postoperative AKI was 40.8% (454/1 112), of which grade 2-3 AKI accounted for 11.9%. Multivariate analysis showed that age (OR=1.049, 95%CI 1.022-1.077, P<0.001), body mass index (OR=1.065, 95%CI 1.010-1.123, P=0.020) and time interval between preoperative coronary angiography and cardiac surgery within 24 hours (OR=1.625, 95%CI 1.116-2.364, P=0.011) were independent predictors of postoperative AKI. Patients who underwent coronary angiography within 24 hours before surgery had a 10.6% higher incidence of postoperative AKI compared to those who underwent angiography ≥24 hours before surgery (P=0.004). Patients who underwent valve surgery with or without coronary artery bypass grafting (CABG) had a higher risk of AKI than those who only underwent CABG. The in-hospital stay of patients who developed AKI was 2 days longer than those without AKI. However, undergoing coronary angiography within 24 hours before cardiac surgery did not prolong the length of ICU stay or hospital stay, nor did it increase the risk of death or renal failure after the operation. Conclusion Undergoing coronary angiography within 24 hours before cardiac surgery increases the risk of postoperative AKI.

Objective:To observe the changes of macular structure and microvessels in eyes with diabetes macular ischemia (DMI).Methods:A retrospective case study. From January 2023 to July 2023, 23 patients of 31 eyes diagnosed with DMI at Tangshan Ophthalmological Hospital were included in this study. Among them, there were 14 males with 23 eyes; Female cases with 8 eyes. Age were (59.5±4.6) years old. According to the DMI grading standard formulated by the research group for early treatment of diabetes retinopathy, the patients were divided into mild DMI group, moderate DMI group, and severe DMI group, with 8, 12, and 11 eyes respectively. The blood flow density (VD), perfusion area (FA), small vessel VD (SVD), inner retinal capillary plexus VD, FA, and outer retinal, choroidal, and ganglion cell complex (GCC) thickness within 1 mm of the macular fovea in retinal superficial vascular plexus (SVP)were measured using a scanning frequency light source optical coherence tomography instrument. The changes in macular structure and microvasculature in the affected eyes of different degrees of DMI groups were compared and observed. Inter group comparisons were conducted using one-way ANOVA or Kruskal Wallis H-test. Spearman correlation analysis was used to analyze the correlation between DMI severity and GCC, outer retina, choroid thickness, VD, FA and SVP VD, SVD and FA in inner retina. Results:The GCC ( F=70.670), outer retinal thickness ( H=12.393), VD ( F=105.506), SVD ( H=25.300), FA ( F=107.655), and VD ( H=24.098) and FA ( H=25.300) of the retinal SVP in the mild, moderate, and severe DMI groups were compared, and the differences were statistically significant ( P<0.05). There was no statistically significant difference in choroidal thickness ( H=2.441, P>0.05). Pairwise comparison between groups: VD, SVD, FA of GCC thickness and SVP, and VD of inner retina were statistically significant between severe DMI group and moderate DMI group, and between moderate DMI group and mild DMI group ( P<0.05). The thickness of outer retina was statistically significant between severe DMI group and moderate DMI group ( P<0.05). Inner retinal FA: there were statistically significant differences between severe DMI group, moderate DMI group and mild DMI group ( P<0.05). The correlation analysis results showed that GCC ( r s=-0.918), outer retinal thickness ( r s=-0.448), and inner retinal VD ( r s=-0.894) and FA ( r s=-0.918), as well as VD ( r s=-0.919), SVD ( r s=-0.924), and FA ( r s=-0.939) of retinal SVP, were all negatively correlated with the degree of DMI ( P<0.05). There was no correlation between choroidal thickness and degree of DMI ( r s=-0.081, P>0.05). Conclusion:The thickness of GCC, outer retina and choroid, the VD, SVD, and FA of the retinal SVP, the VD and FA of inner retina are all reduced in eyes with different degrees of DMI, while all of them are negatively correlated with the degree of DMI, except for choroid thickness.

Objective To investigate the therapeutic effect and underlying mechanism of Qishishenshu Capsule on renal fibrosis in mice with early diabetic nephropathy(DN).Methods A DN mouse model was established by multiple injections of streptozotocin.The mice were randomly divided into a normal group(NC),model group(DN),and Qishi group(QS)(0.9 g/(kg·d)),with eight mice in each group.Mice were gavaged continuously for 4 weeks,and fasting blood glucose(FBG)was measured weekly.Four weeks later,urinary albumin/creatinine(UACR),serum creatinine,and blood urea nitrogen were measured.Hematoxylin-eosin,periodicacid-Schiff,and Sirius red staining were used to analyze renal pathological changes.Real-time fluorescence quantitative reverse-transcription polymerase chain reaction was used to detect the mRNA levels of fibronectin(FN),collagen type Ⅰ alpha 1(Col1a1),and α-smooth muscle actin(α-SMA).Immunohistochemistry and Western blot were performed to detect FN,collagen type Ⅰ(Collagen Ⅰ),collagen typeⅢ(Collagen Ⅲ),α-SMA,Podocin,Nephrin,and transforming growth factor-β1/SMAD family member2/3(TGF-β1/Smad2/3)pathway-related proteins.Results Compared with mice in the NC group,those in the DN group showed significantly higher levels of FBG and UACR(P＜0.001),and mesangial hyperplasia,basement membrane thickening,and collagen deposition in the renal tissue.The mRNA levels of FN,Col1a1,and α-SMA were increased(P＜0.05).Protein levels of Podocin and Nephrin were decreased(P＜0.05).The levels of FN,Collagen I,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were increased(P＜0.05).Compared with the DN group,the QS group's level of UACR was decreased(P＜0.05),their renal pathological injury was alleviated,and mRNA levels of FN,Collagen Ⅰ,andα-SMA were attenuated(P＜0.05);whereas their protein levels of Podocin and Nephrin were elevated(P＜0.05).The levels of FN,Collagen Ⅰ,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were also decreased(P＜0.05).Conclusions Qishishenshu Capsule improved renal fibrosis in DN mice,probably through the inhibition of the TGF-β1/Smad2/3 signaling pathway.

BACKGROUND: : Tourette syndrome (TS) is a neuropsychiatric disorder with onset in childhood that warrants effective therapies. Gut microbiota can affect central physiology and function via the microbiota-gut-brain axis. Therefore, the gut microbiota plays an important role in some mental illnesses. A small clinical trial showed that fecal microbiota transplantation (FMT) may alleviate TS symptoms in children. Herein, FMT effects and mechanisms were explored in a TS mouse model. METHODS: : TS mice model (TSMO) (n = 80) were established with 3,3'-iminodipropionitrile, and 80 mice were used as controls. Mice were grouped into eight groups and were subjected to FMT with feces from children or mice with or without TS, or were given probiotics. Fecal specimens were collected 3 weeks after FMT. 16S rRNA sequencing, behavioral observation, and serum serotonin (5-HT) assay were performed. Differences between groups were analyzed using Mann-Whitney U test and Kolmogorov-Smirnov (KS) tests. RESULTS: : A total of 18 discriminative microbial signatures (linear discriminant analysis score > 3) that varied significantly between TS and healthy mice (CONH) were identified. A significant increase in Turicibacteraceae and Ruminococcaceae in TSMO after FMT was observed (P  < 0.05). Compared with non-transplanted TSMO, the symptoms of those transplanted with feces from CONH were alleviated (W = 336, P = 0.046). In the probiotic and FMT experiments, the serum 5-HT levels significantly increased in TSMO that received probiotics (KS = 1.423, P = 0.035) and in those transplanted with feces from CONH (W = 336.5, P = 0.046) compared with TSMO without transplantation. CONCLUSIONS : This study suggests that FMT may ameliorate TS by promoting 5-HT secretion, and it provides new insights into the underlying mechanisms of FMT as a treatment for TS.
Animals ; Disease Models, Animal ; Fecal Microbiota Transplantation ; Gastrointestinal Microbiome/physiology* ; Mice ; RNA, Ribosomal, 16S/genetics* ; Serotonin ; Tics ; Tourette Syndrome/therapy*

Objective:To analyze locoregional recurrence (LRR) pattern of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, with and without adjuvant radiotherapy (RT). Methods:A total of 5442 eligible patients with breast cancer from 12 Chinese centers were included. The LRR sites and the effect of RT at different sites on recurrence in patients with and without RT were analyzed. The Kaplan-Meier method was used to calculate the cumulative LRR rate, and the difference was compared by the log-rank test.Results:With a median follow-up time of 63.8 months for the entire cohort, 395 patients developed LRR. The chest wall and supraclavicular fossa were the most common LRR sites, regardless of RT or molecular subtypes. The 5-year chest wall recurrence rates for patients with and without chest wall irradiation were 2.5% and 3.8%( P=0.003); the 5-year supraclavicular lymph nodal recurrence rates for patients with and without supraclavicular fossa irradiation were 1.3% and 4.1%( P<0.001); the 5-year axillary recurrence-free rates for patients with and without axillary irradiation were 0.8% and 1.5%( HR=0.31, 95% CI: 0.04-2.23, P=0.219); and the 5-year internal mammary nodal recurrence-free rates for patients with and without internal mammary nodal irradiation were 0.8% and 1.5%( HR=0.45, 95% CI: 0.11-1.90, P=0.268). Conclusions:The chest wall and supraclavicular fossa are the most common LRR sites of patients with pT 1-2N 1 breast cancer after modified radical mastectomy, which is not affected by adjuvant RT or molecular subtypes. The chest wall and supraclavicular fossa irradiation significantly reduce the risk of recurrence in the corresponding area. However, axillary and internal mammary nodal irradiation has no impact on the risk of recurrence in the corresponding area.

Objective:To explore the method of scar management after the operation of congenital syndactyly.Methods:The clinical data of children with congenital cutaneous skin syndactyly of the hand admitted to the Department of Orthopedics of the First Affiliated Hospital of Air Force Medical University from January 2018 to January 2020 were retrospectively analyzed. The procedures of finger dividing were performed under general anesthesia, and the fingers were bandaged in an extended position after the operation. After the incision was healed entirely, anti-scar drugs were used in conjunction with the reconstruction of the webbed with a self-adhesive soft silicone film, wrapped with a self-adhesive elastic bandage for one year, and an extended finger brace was worn for 3 to 6 months. Postoperative complications, finger shape, and appearance were followed up. At the last follow-up, family members of the children were asked to evaluate the reconstruction effect of the webbed shape, scar condition, and flexion and extension of the affected finger.Total active movement( TAM )evaluation criteria were used to evaluate the flexion and extension function of the affected finger. The Vancouver Scar Scale (VSS) was used to evaluate the postoperative cicatrix.Results:A total of 28 children with 32 syndactyly were included, including 12 males and 16 females. The age ranged from 12 to 34 months (mean, 18.3 months). The syndactyly was unilateral in 24 patients (left in 14 and right in 10) and bilateral in 4. Among all the 32 syndactyly, there were 16 cases of the middle-ring finger, 9 cases of the ring-little finger, 5 cases of the index-middle finger, and 2 cases of the thumb-index finger. All the children were followed up for 13~36 months (mean, 28.6 months). No complications such as infection and abnormal blood supply occurred in the early postoperative period. The shape and appearance of the fingers were in good condition. While using a self-adhesive soft silicone membrane, two children developed a local rash, which was improved by desensitization treatment and continued to use usually. The family members were satisfied with the shape of the reconstructed finger web, the scar, and the flexion and extension of the affected finger. TAM evaluation criteria showed a 100% excellent flexion and extension function rate. VSS showed a 100% excellent-good rate of scar status.Conclusions:After incision healing of the reconstruction surgery for pediatric congenital syndactyly, the use of anti-scar drugs, including self-adhesive elastic bandages and split-finger braces, can effectively prevent postoperative scar hyperplasia and better consolidate the treatment effect, with distinct clinical application value.

Objective:To explore the method of scar management after the operation of congenital syndactyly.Methods:The clinical data of children with congenital cutaneous skin syndactyly of the hand admitted to the Department of Orthopedics of the First Affiliated Hospital of Air Force Medical University from January 2018 to January 2020 were retrospectively analyzed. The procedures of finger dividing were performed under general anesthesia, and the fingers were bandaged in an extended position after the operation. After the incision was healed entirely, anti-scar drugs were used in conjunction with the reconstruction of the webbed with a self-adhesive soft silicone film, wrapped with a self-adhesive elastic bandage for one year, and an extended finger brace was worn for 3 to 6 months. Postoperative complications, finger shape, and appearance were followed up. At the last follow-up, family members of the children were asked to evaluate the reconstruction effect of the webbed shape, scar condition, and flexion and extension of the affected finger.Total active movement( TAM )evaluation criteria were used to evaluate the flexion and extension function of the affected finger. The Vancouver Scar Scale (VSS) was used to evaluate the postoperative cicatrix.Results:A total of 28 children with 32 syndactyly were included, including 12 males and 16 females. The age ranged from 12 to 34 months (mean, 18.3 months). The syndactyly was unilateral in 24 patients (left in 14 and right in 10) and bilateral in 4. Among all the 32 syndactyly, there were 16 cases of the middle-ring finger, 9 cases of the ring-little finger, 5 cases of the index-middle finger, and 2 cases of the thumb-index finger. All the children were followed up for 13~36 months (mean, 28.6 months). No complications such as infection and abnormal blood supply occurred in the early postoperative period. The shape and appearance of the fingers were in good condition. While using a self-adhesive soft silicone membrane, two children developed a local rash, which was improved by desensitization treatment and continued to use usually. The family members were satisfied with the shape of the reconstructed finger web, the scar, and the flexion and extension of the affected finger. TAM evaluation criteria showed a 100% excellent flexion and extension function rate. VSS showed a 100% excellent-good rate of scar status.Conclusions:After incision healing of the reconstruction surgery for pediatric congenital syndactyly, the use of anti-scar drugs, including self-adhesive elastic bandages and split-finger braces, can effectively prevent postoperative scar hyperplasia and better consolidate the treatment effect, with distinct clinical application value.

Objective:To explore the method of scar management after the operation of congenital syndactyly.Methods:The clinical data of children with congenital cutaneous skin syndactyly of the hand admitted to the Department of Orthopedics of the First Affiliated Hospital of Air Force Medical University from January 2018 to January 2020 were retrospectively analyzed. The procedures of finger dividing were performed under general anesthesia, and the fingers were bandaged in an extended position after the operation. After the incision was healed entirely, anti-scar drugs were used in conjunction with the reconstruction of the webbed with a self-adhesive soft silicone film, wrapped with a self-adhesive elastic bandage for one year, and an extended finger brace was worn for 3 to 6 months. Postoperative complications, finger shape, and appearance were followed up. At the last follow-up, family members of the children were asked to evaluate the reconstruction effect of the webbed shape, scar condition, and flexion and extension of the affected finger.Total active movement( TAM )evaluation criteria were used to evaluate the flexion and extension function of the affected finger. The Vancouver Scar Scale (VSS) was used to evaluate the postoperative cicatrix.Results:A total of 28 children with 32 syndactyly were included, including 12 males and 16 females. The age ranged from 12 to 34 months (mean, 18.3 months). The syndactyly was unilateral in 24 patients (left in 14 and right in 10) and bilateral in 4. Among all the 32 syndactyly, there were 16 cases of the middle-ring finger, 9 cases of the ring-little finger, 5 cases of the index-middle finger, and 2 cases of the thumb-index finger. All the children were followed up for 13~36 months (mean, 28.6 months). No complications such as infection and abnormal blood supply occurred in the early postoperative period. The shape and appearance of the fingers were in good condition. While using a self-adhesive soft silicone membrane, two children developed a local rash, which was improved by desensitization treatment and continued to use usually. The family members were satisfied with the shape of the reconstructed finger web, the scar, and the flexion and extension of the affected finger. TAM evaluation criteria showed a 100% excellent flexion and extension function rate. VSS showed a 100% excellent-good rate of scar status.Conclusions:After incision healing of the reconstruction surgery for pediatric congenital syndactyly, the use of anti-scar drugs, including self-adhesive elastic bandages and split-finger braces, can effectively prevent postoperative scar hyperplasia and better consolidate the treatment effect, with distinct clinical application value.