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 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 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 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.

Objective:To investigate the application and surgical effect of lobed anterolateral femoral chimeric perforator flap in repairing the complex soft tissue defects of limbs.Methods:Clinical materials of 52 cases with complex soft tissue defects of limbs were retrospectively reviewed from October 2013 to October 2020. Thirty-two males and 20 females were reviewed, and the age of them ranged from 6 to 65(mean 39) years old. The injuries include 17 cases of traffic accidents, 15 machine crush injuries, 12 engineering accidents, and 8 extensive resection of malignant tumors. The size of soft tissue defects ranged from 26.0 cm×10.0 cm to 35.0 cm×15.0 cm. Lobed anterolateral thigh chimeric perforator flaps or lobed Flow-through chimeric perforator flaps were designed according to the soft tissue defects and blood circulation of the distal extremities. Start from 8-month post-operation, upper limb function was assessed according to the Upper Limb Function Assessment Standard of Hand Surgery Branch of Chinese Medical Association and Disability of arm shoulder and hand(DASH) score scale, and Fugl-Meyer score system was used to evaluate the lower limb function. Long-term follow-up was conducted through WeChat video call and outpatient clinic evaluation.Results:All flaps survived. One patient with crush injury got delayed wound healing due to deep hemaetoma after surgery, and the rest were healed at the first stage. All the cases entered followed-up, ranged from 6 to 20(mean 8) months. The appearance of the flaps was satisfactory, and the color of the flaps has no significant difference compared with the recipient area. No recurrence of tumor was reported during the follow-up period. According to the Upper Limb Function Assessment Standard of Hand Surgery Branch of Chinese Medical Association, there were 21 excellent cases and 8 good cases, and the total excellent and good rate was 100%. The DASH score of upper limbs ranges from 11.5 to 45.1(mean 25.4±13.7), and the Fugl-Meyer score of lower limbs range from 18.6 to 31.8(mean 26.2±11.2) at the last follow-up.Conclusion:The complex soft tissue defect of limbs is a common complication caused by high-energy trauma, which is difficult to repair. The lobed anterolateral thigh chimeric perforator flap through a Decompose-Combine and arrange method for reconstruction, which is flexible in flap design. A variety of tissue flaps can be obtained by anastomosing only one group of blood vessels. It is an ideal choice for repairment of complex soft tissue defects of limbs.

Objective:To observe the multimodal imaging characteristics of tamoxifen retinopathy.Methods:A retrospective case study. From January 2019 to December 2021, 4 patients (8 eyes) with tamoxifen retinopathy diagnosed in Tangshan Eye Hospital were included in the study. All patients were female, with sick binoculus. The age was 59.5±4.6 years. After breast cancer resection, tamoxifen 20 mg/d was taken orally consecutively, including 1, 1, and 2 cases who took tamoxifen orally for 5, 7, and ≥10 years. All eyes were examined by fundus color photography, optical coherence tomography (OCT), OCT angiography (OCTA), fundus fluorescein angiography (FFA), and fundus autofluorescence (AF). The multi-mode image features of the fundus of the affected eyes were observed.Results:The yellow white dot crystal like material deposition in the macular area was observed in all eyes. In fundus AF examination, macular area showed patchy strong AF. FFA examination showed telangiectasia and fluorescein leakage in macular area at late stage. OCT showed that punctate strong reflexes could be seen between the neuroepithelial layers in the macular region with the formation of a space between the neuroepithelial layers, the interruption of the elliptical zone (EZ), and the formation of a hole in the outer lamella including 4, 5 and 3 eyes; The thickness of ganglion cells in macular region decreased in 7 eyes. OCTA showed that the blood flow density of the superficial retinal capillary plexus around the arch ring was decreased, and the retinal venules were dilated in 2 eyes; Deep capillary plexus (DCP) showed telangiectasia.Conclusion:Deposition of yellowish white dot like crystals can be seen in the macular region of tamoxifen retinopathy; dotted strong reflex between neuroepithelial layers, cavity formation, thinning of ganglion cell layer, EZ middle fissure and outer lamellar fissure; DCP capillaries and venules around the arch were dilated; telangiectasia in macular region; flaky strong AF in macular region.

Multiple plant lineages have independently evolved sex chromosomes and variable kary-otypes to maintain their sessile lifestyles through constant biological innovation.Morus notabilis,a dioecious mulberry species,has the fewest chromosomes among Morus spp.,but the genetic basis of sex determination and karyotype evolution in this species has not been identified.In this study,three high-quality genome assemblies were generated for Morus spp.[including dioecious M.notabilis(male and female)and Morus yunnanensis(female)]with genome sizes of 301-329 Mb and were grouped into six pseudochromosomes.Using a combination of genomic approaches,we found that the putative ancestral karyotype of Morus species was close to 14 protochromosomes,and that sev-eral chromosome fusion events resulted in descending dysploidy(2n=2x=12).We also charac-terized a～6.2-Mb sex-determining region on chromosome 3.Four potential male-specific genes,a partially duplicated DNA helicase gene(named MSDH)and three Ty3_Gypsy long terminal repeat retrotransposons(named MSTG1/2/3),were identified in the Y-linked area and considered to be strong candidate genes for sex determination or differentiation.Population genomic analysis showed that Guangdong accessions in China were genetically similar to Japanese accessions of mul-berry.In addition,genomic areas containing selective sweeps that distinguish domesticated mul-berry from wild populations in terms of flowering and disease resistance were identified.Our study provides an important genetic resource for sex identification research and molecular breeding in mulberry.

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.