Objective:To investigate the mechanism of curcumin affecting HIV-1 infection co-receptor CCR5 by regulating transcription factor FOXP3.Methods:Binding sites of transcription factor FOXP3 on CCR5 promoter were predicted and analyzed by bioinformatics method.AutoDock 4.2 software was used to connect curcumin and FOXP3 flexibly.MTT assay was used to detect cyto-toxcity of curcumin on activity of Jurkat cells.qRT-PCR and Western blot were used to detect expression levels of CCR5 and FOXP3 mRNA and protein in Jurkat cells that were treated with different concentrations of curcumin.pcDNA3.1-FOXP3 expression vector was built and combined with the prediction results of transcription factors.The mutant CCR5 gene fragment was amplified by Overlap PCR,and the mutant CCR5 promoter recombinant vector pFireRluc-Mt-CCR5 was constructed.Binding site between transcription fac-tor FOXP3 and CCR5 promoter was verified by double luciferase reporter gene assay.Results:Results of JASPAR transcription factor prediction showed that there was a binding site between CCR5 promoter and transcription factor FOXP3;molecular docking results showed that curcumin could bind to the active region of FOXP3;MTT results showed that curcumin inhibited the activity of Jurkat cells after 24 hours,and the IC50 was 34.48 μmol/L.qRT-PCR and Western blot showed that expression levels of CCR5 and FOXP3 mRNA and protein were decreased in a dose-dependent manner after different concentrations of curcumin treated Jurkat cells;double luciferase reporter gene confirmed that FOXP3 could bind to CCR5 promoter,and the transcription factor FOXP3 could regulate the activity of CCR5 promoter;results of the recovery experiment of FOXP3 on curcumin showed that when the curcumin concentration was 60 μmol/L,relative value of luciferase activity in HEK293T cells with pcDNA3.1-FOXP3 and pFireRluc-Wt-CCR5 was signifi-cantly higher than that in pFireRluc-Wt-CCR5+curcumin-60 group(P<0.01).Conclusion:FOXP3 can regulate the activity of CCR5 promoter,and the mechanism may be that curcumin affects activity of CCR5 promoter by acting on binding site of FOXP3 and CCR5 promoter.

Objective:To investigate the effect of perineal single-port robot-assisted radical prostatectomy.Methods:A retrospective analysis was conducted on clinical data from 60 patients who underwent perineal single-port robot-assisted laparoscopic radical prostatectomy at our hospital between July 2019 and July 2022. The mean age of the patients was (65.9±7.6) years and the mean BMI was (24.1±2.9) kg/m 2. The median (IQR) prostate volume was 32.7 (23.8, 41.2) ml, and the median (IQR) preoperative PSA value was 8.8 (6.8, 12.6) ng/ml. Preoperative pathology revealed a Gleason score of 6 in 21 patients, Gleason score of 7 in 35 patients and Gleason score of 8 in 4 patients. There were 12 patients clinically staged as T 1 and 48 patients as T 2. A total of 18 patients underwent a total of 23 previous abdominopelvic surgeries. The patient is placed in an exaggerated lithotomy position with the head down and feet elevated approximately 15°. A 3-5 cm incision was made approximately 2 cm above on the mid-perineum between the bilateral ischial tuberosities. Next, the rectourethral muscle was divided, and the space anterior to the rectum was developed by blunt dissection. The levator ani muscles were separated to expose Denonvilliers’ fascia. Then, the disposable multi-channel laparoscopic surgical access system is inserted with a surgical wound protector. Denonvilliers’ fascia was incised transversely and the ampulla of the vas deferens, which were subsequently divided. Blunt separation is performed on both sides along the capsule of the prostate, and then, the vascular pedicles of the prostate are ligated. The membranous urethra was severed after complete urethral separation at the tip of the prostate at the urethral junction. The bladder neck was freed and dissected. The prostate and seminal vesicles were removed and a vesicourethral anastomosis is performed. A perineal drain were left in place. Preoperative and postoperative variables, complications, early urinary continence rate(Return of urinary continence status was defined as using no more than one safety pad per day) and oncological outcomes of patients were recorded. Results:All 60 surgeries were successfully completed without conversions or additional incisions. The median (IQR) total operative time was 200.0(153.8, 236.3) min, the median (IQR) console operating time was 107.5(90.0, 150.0) min and the median (IQR) estimated blood loss was 50.0(50.0, 100.0) ml. Positive surgical margins were detected in five patients (8.3%). The continence rate was 43.1%(22/51), 64.7%(33/51), 92.0%(46/50) and 98.0%(49/50), and the PSA undetectable rate was 94.6%(48/51), 98.2%(49/51), 96.6%(47/50) and 100%(50/50) at the 1, 3, 6, and 12 months after surgery. Only 1(1.7%) patient experienced biochemical recurrence 9 months after surgery. The overall complication rate was 20%, including two cases of acute respiratory distress syndrome, one case of rectal injury, one case of urinary tract injury, two cases of poor wound healing, three cases of incision infection, two cases of urinary tract infection and one case of bladder neck-urethral orifice anastomotic stricture.Conclusions:Perineal single-port robot-assisted radical prostatectomy might be safe and feasible surgical treatments for localized prostate cancer, especially for patients with a history of complex abdominal or pelvic surgery. It also showed advantages in early continence. The anatomical structure of the perineal region should be considered, and the correct incision position should be chosen. Specific incision protection measures should also be used for the incision in this particular area of the perineal region to reduce the risk of perioperative complications.

Objective To establish preoperative and postoperative femoral-pelvic-lumbar spine models of patients with developmental dysplasia of the hip(DDH)and healthy volunteers and to study the biomechanical effects of curved periacetabular osteotomy on the lumbar spine.Methods Preoperative and postoperative femoral-pelvic-lumbar spine DICOM data from four patients with DDH and one healthy volunteer were acquired using CT scanning technology,and a three-dimensional finite element model was constructed.The offset method was used to divide the cortical and cancellous bones in Geomagic and the lumbar cartilage,sacroiliac joint,pubic symphysis,and other cartilages were added to SolidWorks.The model was analyzed using ANSYS for finite element analysis,and the gait was the mid-stage of single-leg support during slow walking.The biomechanical changes in the lumbar spine of patients with DDH before and after surgery were analyzed and compared,and the biomechanical data of the lumbar spine of patients after surgery were compared with those of healthy volunteers.Results The femoral-pelvic-lumbar spine models of four patients and a healthy volunteer were established.The results obtained by the established models under each working condition were within the range of the referenced literature,and the validity of the models was proved.The postoperative stresses on the lumbar spine,femoral neck,annulus fibrosus,and nucleus pulposus were much smaller than those of the patients in the preoperative state,and the postoperative stresses on the lumbar spine,femoral neck,annulus fibrosus,and nucleus pulposus of the patients were similar to those of healthy volunteers.Conclusions Curved periacetabular osteotomy significantly reduced the stresses on the lumbar spine and intervertebral discs.Additionally,the stresses on the annulus fibrosus were more uniform after surgery,which indicated that curved periacetabular osteotomy will adjust patients to a healthy state.This study provides a biomechanical basis for the clinical treatment of DDH and helps optimize surgical plans.

BACKGROUND:Kidney deficiency is the main pathogenesis of osteoporosis.To study the relationship between the two major syndrome types of kidney deficiency,Kidney-Yang deficiency and Kidney-Yin deficiency,is beneficial for the development of clinical diagnosis and treatments based on the combination of disease and syndrome. OBJECTIVE:To evaluate the biomechanical differences of the rat femurs with Kidney-Yang deficiency and Kidney-Yin deficiency caused by Yougui pills,and to demonstrate the scientific efficacy of medication based on the combination of disease and syndrome in osteoporosis from a biomechanical perspective. METHODS:The bilateral ovaries of 60 female Sprague-Dawley rats were surgically removed to establish an ovariectomized osteoporosis model.At 10 weeks after modeling,all the rats were randomly divided into a Kidney-Yang deficiency group(n=30)and a Kidney-Yin deficiency group(n=30).Rats with Kidney-Yang deficiency were given gluteal intramuscular injection of hydrocortisone,while rats with Kidney-Yin deficiency were orally administered with thyroid tablet suspension,once a day,for 14 consecutive days.After successful modeling,20 rats in each group were given a suspension of Yougui pills by gavage once a day for 12 consecutive weeks and the remaining 10 rats were used as the control group without intervention.After gavage,the microstructural parameters of the bone were measured using Micro-CT scanning.Three-point bending,finite element simulation,femoral head compression,and surface indentation distribution experiments of the femurs were performed on a mechanical testing machine. RESULTS AND CONCLUSION:Micro-CT revealed that the femoral bone density,bone volume fraction,bone surface density,trabecular number,and trabecular separation were improved in the Kidney-Yin deficiency+Yougui pills group compared with the Kidney-Yin deficiency group(P＜0.05);the femoral bone volume fraction,bone surface density,trabecular number,and trabecular thickness were improved in the Kidney-Yang deficiency+Yougui pills group compared with the Kidney-Yang deficiency group(P＜0.05).The three-point bending experiment showed that the femur elastic modulus,maximum bending strength and bending fracture strength were decreased(P＜0.05)and toughness was increased(P＜0.05)in the Kidney-Yang deficiency+Yougui pills group compared with the Kidney-Yang deficiency group.Finite element simulation showed that Yougui pills could significantly improve the bending resistance of the femurs in the Kidney-Yang deficiency group,but had no significant effect on the Kidney-Yin deficiency group.The femoral head compression experiments showed that Yougui pills could enhance the ability of the femoral head to resist deformation in the Kidney-Yang deficiency group,but there was no significant difference in the effect of Yougui pills on the surface properties of the femoral head in the Kidney-Yin deficiency group and the Kidney-Yang deficiency group.To conclude,Yougui pills can significantly enhance the biomechanical properties of the osteoporotic bones with Kidney-Yang deficiency,but have no significant effect on the osteoporotic bone with Kidney-Yin deficiency.

BACKGROUND:Previous studies have successfully constructed erythropoietin-overexpressed umbilical cord mesenchymal stem cells.It was found that the apoptosis of ischemic and hypoxic human neuroblastoma cell line(SH-SY5Y)was significantly reduced by erythropoietin-overexpressed umbilical cord mesenchymal stem cells. OBJECTIVE:To explore the possible neuroprotective mechanisms of erythropoietin-overexpressed umbilical cord mesenchymal stem cells against ischemic-hypoxic SH-SY5Y and their associated epigenetic mechanisms. METHODS:Oxygen-glucose deprivation was applied to ischemia-hypoxia-induced SH-SY5Y cell injury,and multifactorial assays were applied to detect the expression levels of inflammatory factors in the cells before and after hypoxia and co-culture,respectively,with mesenchymal stem cells,as well as lentiviral-transfected null-loaded plasmids of the negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression levels of supernatant inflammatory factors were detected by multifactor assay after co-culture.Proteomics was used to detect the differentially expressed proteins of negative control mesenchymal stem cells and erythropoietin-overexpressed umbilical cord mesenchymal stem cells.Cleavage under targets and tagmentation sequencing was applied to detect genomic H3K4me2 modification,and joint analysis was conducted with RNA-sequencing.Lentiviral vector infection was applied to construct the stable knockdown of REST in SH-SY5Y cells.qRT-PCR and western blot assay were performed to detect the expression level of REST.The apoptosis was detected by flow cytometry after co-culture of oxygen-glucose deprivation treatment with erythropoietin-overexpressed umbilical cord mesenchymal stem cells.The expression difference of H3K36me3 group proteins was detected by western blot assay,and transcriptome sequencing was performed to analyze the differentially expressed genes. RESULTS AND CONCLUSION:(1)Compared with the control group,monocyte chemotactic protein 1,interleukin-6,interleukin-18,and interleukin-1 beta,interferon α2,and interleukin-23 levels significantly increased in the cerebrospinal fluid supernatant of patients with ischemic-hypoxic encephalopathy(P<0.01).(2)After co-culturing SH-SY5Y cells with erythropoietin-overexpressed umbilical cord mesenchymal stem cells under ischemia and hypoxia,the expression levels of monocyte chemotactic protein 1 and interleukin-6 were significantly reduced.(3)Analysis of protein network interactions revealed significant downregulation of monocyte chemotactic protein 1,interleukin-6 related regulatory proteins CXCL1 and BGN.(4)Transcriptome sequencing analysis found that pro-inflammatory genes were down-regulated,and functional enrichment of histone modifications,and the expression of transcription factors REST and TET3 significantly up-regulated in the erythropoietin-overexpressed umbilical cord mesenchymal stem cell group compared with the negative control mesenchymal stem cell group.(5)Combined analysis of transcriptome sequencing and cleavage under targets and tagmentation revealed changes in epigenetic levels as well as significant activation of the promoter regions of transcription factors REST and TET3.(6)Stable knockdown REST in SH-SY5Y cells was successfully constructed;the transcript levels of REST mRNA and protein expression were both decreased.(7)After the REST knockdown SH-SY5Y cells were co-cultured with erythropoietin-overexpressed umbilical cord mesenchymal stem cells,apoptosis was significantly increased and H3K36me3 expression was significantly decreased.Transcriptome sequencing results showed that the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2 was reduced at mRNA transcription level(P<0.01).(8)It is concluded that erythropoietin-overexpressed umbilical cord mesenchymal stem cells activated the expression of REST and TET3 by altering the kurtosis of H3K4me2 and upregulated the modification level of H3K36me3,which in turn regulated the expression of inflammation-related genes Aldh1l2 and Cth,as well as apoptosis-suppressor genes Mapk8ip1 and Sod2,and facilitated neuronal survival.

【Objective】 To investigate the status quo of disease perception and partner support of erectile dysfunction (ED) patients, and to analyze their correlation. 【Methods】 With convenient sampling method, 220 ED patients in a first-class hospital in Taiyuan were surveyed with the short version of disease perception questionnaire (BIPQ) and partner support coping questionnaire (DCI). 【Results】 The BIPQ score was (41.90±7.33), and the DCI score was (116.79±20.37). Pearson correlation analysis showed that except life influence, treatment control and emotional influence, the mutual support dimension of partner support of ED patients was correlated with all dimensions of disease perception (r=-0.173, 0.151, -0.182, 0.163, 0.188,P<0.05).Except cognitive comprehension, the negative support dimension of partner support of ED patients was correlated with the dimensions and total score of disease perception (r=0.399, 0.185, -0.167, -0.306, 0.269, 0.445, 0.320, 0.357,P<0.05). 【Conclusion】 ED patients have negative disease perception, and their partners have better coping strategies. Medical staff may pay more attention to partner support so as to reduce patients’ negative perception of disease and promote recovery.

The source and process of mandible development are significantly different from those of other bones in the body,and abnormal development can lead to various bone-related diseases,seriously affecting the quality of life of patients.In recent years,the role of the Hedgehog signaling pathway in bone development has received increasing atten-tion.The Hedgehog gene includes three subtypes:Sonic Hedgehog(Shh),Indian Hedgehog(Ihh),and Desert Hedgehog(Dhh).Shh and Ihh can participate in bone metabolism regulation through various pathways,with Shh primarily involved in limb development and Ihh playing a key role in endochondral osteogenesis.The Hedgehog signaling pathway includes Hedgehog signaling protein ligands,Patched(Ptch)receptors,Smoothed(Smo)receptors,nuclear transcription factors,glioma-associated oncogene homologues(Gli),and downstream target genes.The activation of typical Hedgehog signal-ing pathways requires the involvement of Gli,whereas atypical Hedgehog signaling is mainly regulated by Ptch,Smo,and others.Shh regulates various biological behaviors during early vertebrate embryogenesis,such as organ differentia-tion,neural stem formation,stem cell differentiation and proliferation,limb bone development,and tooth germ develop-ment.During the process of bone cell differentiation,Shh,Ptch1,and Gli1 are expressed in osteoblasts,further promot-ing the differentiation of bone marrow mesenchymal stem cells into osteoblasts and chondrocytes.IHh plays an indis-pensable functional role in bone growth,development,and homeostasis and participates in the formation of intramem-brane bone collars,proliferation,and maturation of chondrocytes.IHh is expressed in mature skull osteoblasts and can act as a promoter of bone factor regulation of Ptch and bone morphogenetic protein(BMP)expression to induce intra-membrane ossification.Brain and muscle ARNT-like protein 1(BMAL1)can regulate the Hedgehog signaling pathway by binding to Ptch1 and Ihh,playing a crucial role in cartilage formation and endochondral osteogenesis in the temporo-mandibular joint.Hedgehog signal activators can improve the reduction in mandibular bone mass caused by BMAL1 de-ficiency.Hedgehog signaling imbalance can have a significant impact on bone development and lead to a series of bone diseases,such as abnormal bone development,fractures,osteoporosis,and osteoarthritis.The mechanism of the Hedge-hog signaling pathway in relation to mandibular diseases has not been fully elucidated,and future research should seek to further explore Hedgehog signaling as a potential target for treating mandibular developmental-related diseases.

To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.

Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.