Pathogenic microorganisms are direct causative agents of zoonotic infectious diseases,po-sing severe threats to the livestock industry by inducing massive animal mortality,economic losses in livestock products,and significant risks to human health.The CRISPR/Cas system has been widely adopted in nucleic acid detection of pathogenic microorganisms due to its unique trans-cleavage activity.By leveraging the superior optical properties of nanomaterials,researchers have integrated them with CRISPR/Cas systems to develop numerous visual biosensors,which not only significantly enhance signal output but also substantially reduce detection time and cost.This re-view focuses on five nanomaterials-graphene oxide(GO),gold nanoparticles(AuNPs),MoS2 nanosheets,metal-organic frameworks(MOFs),and quantum dots(QDs)—that have been exten-sively integrated with CRISPR/Cas systems in recent years.We systematically summarize their distinct physical characteristics and specific applications in CRISPR/Cas-based pathogen detection,followed by a concise comparison of the advantages and limitations of different methodologies.Fi-nally,we discuss the prospects for nanomaterials in CRISPR/Cas detection systems,aiming to pro-vide a valuable reference for advancing molecular diagnostics of pathogenic microorganisms.

Objective To explore the medium-and long-term efficacy of modified approach arthroscopy in the treatment of ischial tuberosity cysts in middle-aged and elderly people.Methods Clinical data of 19 cases of ischial tuberosity cyst treated arthroscopically from January 2018 to December 2020 were retrospectively analyzed.The cyst wall was removed by using the central approach of the ischial tuberosity cyst and the 3-5 cm approach of the cyst to the outside of the buttocks.Results The operation time was 30-45 min(mean,36.2±4.8 min).The Visual Analogue Scale(VAS)scores of the pain on the 1st,7th,and 28th d after surgery were 2-6(mean,3.7±1.1),1-4(mean,1.9±0.9),and 0-1(mean,0.5±0.2),respectively.There were 18 cases of incision grade A healing and 1 case of grade B healing.The total hospitalization time was 3-6 d(mean,3.8±1.1 d).There were no complications such as incision infection and subcutaneous hematoma.The 19 cases were followed up for 40-81 months(mean,60.5±10.2 months),and 1 case had cyst recurrence at 3 months after surgery.Conclusion The modified approach arthroscopy is safe and effective in the treatment of ischial tuberosity cysts,having satisfactory medium-and long-term efficacy.

Objective To explore the medium-and long-term efficacy of modified approach arthroscopy in the treatment of ischial tuberosity cysts in middle-aged and elderly people.Methods Clinical data of 19 cases of ischial tuberosity cyst treated arthroscopically from January 2018 to December 2020 were retrospectively analyzed.The cyst wall was removed by using the central approach of the ischial tuberosity cyst and the 3-5 cm approach of the cyst to the outside of the buttocks.Results The operation time was 30-45 min(mean,36.2±4.8 min).The Visual Analogue Scale(VAS)scores of the pain on the 1st,7th,and 28th d after surgery were 2-6(mean,3.7±1.1),1-4(mean,1.9±0.9),and 0-1(mean,0.5±0.2),respectively.There were 18 cases of incision grade A healing and 1 case of grade B healing.The total hospitalization time was 3-6 d(mean,3.8±1.1 d).There were no complications such as incision infection and subcutaneous hematoma.The 19 cases were followed up for 40-81 months(mean,60.5±10.2 months),and 1 case had cyst recurrence at 3 months after surgery.Conclusion The modified approach arthroscopy is safe and effective in the treatment of ischial tuberosity cysts,having satisfactory medium-and long-term efficacy.

Pathogenic microorganisms are direct causative agents of zoonotic infectious diseases,po-sing severe threats to the livestock industry by inducing massive animal mortality,economic losses in livestock products,and significant risks to human health.The CRISPR/Cas system has been widely adopted in nucleic acid detection of pathogenic microorganisms due to its unique trans-cleavage activity.By leveraging the superior optical properties of nanomaterials,researchers have integrated them with CRISPR/Cas systems to develop numerous visual biosensors,which not only significantly enhance signal output but also substantially reduce detection time and cost.This re-view focuses on five nanomaterials-graphene oxide(GO),gold nanoparticles(AuNPs),MoS2 nanosheets,metal-organic frameworks(MOFs),and quantum dots(QDs)—that have been exten-sively integrated with CRISPR/Cas systems in recent years.We systematically summarize their distinct physical characteristics and specific applications in CRISPR/Cas-based pathogen detection,followed by a concise comparison of the advantages and limitations of different methodologies.Fi-nally,we discuss the prospects for nanomaterials in CRISPR/Cas detection systems,aiming to pro-vide a valuable reference for advancing molecular diagnostics of pathogenic microorganisms.

Objective To investigate the clinical efficacy of modified arthroscopic revision release for patients who have gluteal muscle contracture and who have poor outcomes after traditional open surgery.Methods The data of patients who underwent modified arthroscopic revision release for residual symptoms of gluteal muscle contracture after traditional open surgery were retrospectively collected and analyzed.All subjects underwent the procedure between December 2015 and December 2022.The surgical efficacy was assessed by evaluating improvements in specific symptoms,including bilateral lower extremity inequality,hip internal rotation and adduction mobility,squatting with both knees pressed together,and the ability to cross one's legs in supine position,as well as the preoperative and postoperative results for the gluteal muscle contracture functionality scale.Paired t-test was performed to examine whether the differences between preoperative and postoperative measurements were statistically significant.Results A total of 36 patients were followed up systematically,with the mean follow-up period being(22.4±4.9)months.All patients had significantly higher scores for assessment with the gluteal muscle contracture functionality scale at the last follow-up than their preoperative assessment results,showing an increase from the preoperative scores of 40.2±5.5 to 78.4±4.9(P＜0.05).At the follow-up,all patients showed improvement in hip adduction and internal rotation mobility compared with their preoperative status and all patients were able to squat with both knees pressed together.Moreover,only 1 patient still had difficulty in crossing his legs.A total of 27 cases(75%)had preoperative leg length inequality,all of which improved to varying degrees at follow-up.Among all the patients(72 hips/cases),8 cases had subcutaneous hematomas and incisional ecchymosis,which were resolved after conservative treatments such as hot compresses.3 cases showed decreased hip abductor strength,but the muscle strength gradually recovered after postoperative exercise and rehabilitation.There were no complications such as subcutaneous exudate,neurovascular injury,or surgical site infection.Conclusion Modified arthroscopic revision release of gluteus muscle contracture is suitable for cases with poor outcomes after conventional open surgery.

Background::Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer associated with poor prognosis and limited treatment options. The androgen receptor (AR) has emerged as a potential therapeutic target for luminal androgen receptor (LAR) TNBC. However, multiple studies have claimed that anti-androgen therapy for AR-positive TNBC only has limited clinical benefits. This study aimed to investigate the role of AR in TNBC and its detailed mechanism.Methods::Immunohistochemistry and TNBC tissue sections were applied to investigate AR and nectin cell adhesion molecule 4 (NECTIN4) expression in TNBC tissues. Then, in vitro and in vivo assays were used to explore the function of AR and estrogen receptor beta (ERβ) in TNBC. Chromatin immunoprecipitation sequencing (ChIP-seq), co-immunoprecipitation (co-IP), molecular docking method, and luciferase reporter assay were performed to identify key molecules that affect the function of AR. Results::Based on the TNBC tissue array analysis, we revealed that ERβ and AR were positive in 21.92% (32/146) and 24.66% (36/146) of 146 TNBC samples, respectively, and about 13.70% (20/146) of TNBC patients were ERβ positive and AR positive. We further demonstrated the pro-tumoral effects of AR on TNBC cells, however, the oncogenic biology was significantly suppressed when ERβ transfection in LAR TNBC cell lines but not in AR-negative TNBC. Mechanistically, we identified that NECTIN4 promoter –42 bp to –28 bp was an AR response element, and that ERβ interacted with AR thus impeding the AR-mediated NECTIN4 transcription which promoted epithelial–mesenchymal transition in tumor progression. Conclusions::This study suggests that ERβ functions as a suppressor mediating the effect of AR in TNBC prognosis and cell proliferation. Therefore, our current research facilitates a better understanding of the role and mechanisms of AR in TNBC carcinogenesis.

Objective:To observe any effect of electroacupuncture (EA) on the expression of phosphorylated extracellular signal-regulated protein kinase (p-ERK1/2) and phosphorylated cyclic adenosine monophosphate response element binding protein (p-CREB) in the spinal dorsal horns of diabetics experiencing neuropathic pain.Methods:Eight rats were randomly selected from 30 healthy male Sprague-Dawley rats as the normal group (N), and the remaining twenty-two rats were treated with a single high-dose intraperitoneal injection of streptozotocin (STZ) to establish a neuropathic pain model. The rats modeled successfully were randomly divided into a model group (M, n=8) and an EA group ( n=8). In the EA group, electroacupuncture was applied at the bilateral Hou san li and Kunlun acupoints starting on the 15th day after the STZ injection. The daily sessions lasted 30 minutes for 1 week. Body weight (BW), fasting blood glucose (FBG) and paw withdrawal latency (PWL) were observed before the STZ injection and on the 7th, 14th, and 21st days afterward. The expression of p-ERK1/2 and p-CREB in the dorsal horns of the rats′ spinal cords was detected using western blotting. The count of p-CREB-positive cells in the dorsal horns and their co-localization with neurons was detected using immunofluorescence. Results:In comparison with the N group, the average BW of the M group on the 7th, 14th and 21st days after the STZ injection was significantly lower, while the average FBG was significantly higher. There was no significant difference between the M and N groups in the average PWL on the 7th day after the STZ injection, but it had decreased significantly in the M group on the 14th and 21st days. Compared with the M group, the average PWL of the EA group was significantly longer on the 21st day after the injection. The expression of p-ERK1/2 and p-CREB protein in the spines of the M group was significantly higher than in the N group. p-CREB positive cells were more numerous in the M group compared with the N group, while in the EA group they were fewer. P-CREB was co-located with neurons in the spinal dorsal horn.Conclusion:EA can alleviate neuropathic pain effectively, perhaps by inhibiting the expression of p-ERK1/2 and p-CREB in the dorsal horns of the spinal cord.

OBJECTIVETo investigate the effects of microRNA(miRNA)-29b on the proliferation and migration of breast cancer cells and its molecular mechanism.

METHODSThe recombinant lentiviral expression vector (lenti-miRNA-29b) was constructed and transfected into 293T cells to obtain lentivirus particles that were used to infect breast cancer MCF-7 cells. Transfection efficiency of lenti-miRNA-29b in MCF-7 cells was identified by the expression of green fluorescent protein (GFP). The expression of miRNA-29b was detected by real-time PCR. The cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The bioinformatics softwares were used to predict and screen the downstream target genes regulated by miRNA-29b, which were verified by double luciferase reporter gene assay, RT-PCR and Western blot. The effects of screened target gene RTKN on the growth and migration of MCF-7 cells were verified by RTKN siRNA.

RESULTSRecombinant lentiviral expression vector of miRNA-29b were successfully constructed. About 90% and 60% of the breast cancer cells showed green fluorescence in lenti-miRNA-29b and lenti-miRNA-NC groups, respectively. The expression of miRNA-29b in lenti-miRNA-29b group increased significantly compared with the lenti-miRNA-NC group and blank control group (all<0.05); the proliferation and migration ability of MCF-7 cells significantly reduced compared with the control group (all<0.05). The screening with bioinformatics softwares found that the 3'UTR coding region RTKN had the binding site to miRNA-29b; the dual luciferase reporter gene assay showed that the luciferase activity decreased significantly after the MCF-7 cells were co-transfected with wild type RTKN-WT-3'UTR and miRNA-29b mimics report gene vector (<0.05). The RTKN proteins in MCF-7 cells were significantly decreased after transfection with siRNA-RTKN, and the proliferation and migration ability of MCF-7 cells were significantly reduced (all<0.05).

CONCLUSIONSMiRNA-29b can inhibit the proliferation, invasion and metastasis of breast cancer cells by inhibiting the expression of RTKN.

Objective: To investigate the clinical and genetic character of Chinese children with the aarF domain containing kinase 4 (ADCK4)-associated glomerulopathy. Methods: Applying next generation sequencing to detect possible gene mutation(renal disease associated monogene was pooled as one panel) in 69 children with steroid-resistant nephrotic syndrome (SRNS) or persistent proteinuria of unknown origin. Sanger sequencing was used to confirm the significant mutations found in the children and to validate these mutation sites in their patients. Using online software (PolyPhen2, SIFT, Mutation Taster) to predict whether the detected missense mutations were disease causing or not. Collecting and analyzing clinical data of children with ADCK4-associated glomerulopathy, which included onset age, clinical manifestation, and renal pathology. Results: The ADCK4 gene mutation was detected in 8 children with a positive rate of 11.6% (8 out of 69), among which 3 patients carried homozygous c.748G>C mutation, 3 patients carried homologous c.737G>A mutation, 1 patient carried compound heterozygous mutation(c.748G>C and c.737G>A), and 1 patient carried compound heterologous mutation(c.551A>G and c.737G>A). Collectively, there were only 3 mutation sites found in total 8 patients, in which the mutation sites of c.748G>C and c.737G>A had high detection frequency in these 8 patients. These 3 mutation sites were all missense mutation which were predicted to be disease causing by online software and not reported before. The average onset age was 6.5 years (2 years-11.75 years). Four patients presented with SRNS and the other 4 presented with persistent proteinuria. All 8 patients had no extrarenal manifestation, renal biopsy revealed focal segmental glomerulosclerosis (FSGS) in most patients, among which 3 cases had gone to end-stage renal disease (ESRD) at disease onset, and 2 cases progressed to ESRD 2 and 5 years after onset respectively. Seven patients had received glucocorticoid and/or immunosuppressive drug while only one patient getting partial response. All 8 patients were treated with large amount of coenzyme Q₁₀ (15 mg·kg^-1·d^-1) after definite diagnosis of ADCK4 mutation-some patients had acquired encouraging curative effect. Conclusions ADCK4-associated glomerulopathy is not rare especially in the children with SRNS. The onset age is relatively old and the extrarenal manifestation is less common. FSGS is a main pathology type. Patients usually have no response to immunosuppressive therapy, but may benefit from addition of large amount of coenzyme Q₁₀. Some patients may only manifest with insidious proteinuria, causing the early diagnosis to be difficult, which deserves more attention. Three new missense mutations expand disease causing mutation repertoire of ADCK4 gene, among which the two sites of c.748G>C and c.737G>A may be mutation hotspot of ADCK4-associated glomerulopathy in Chinese population, and need further study.

Objective Genetic variants in microRNA (miRNA) binding regions of the gene 3′untranslated region (3′UTR) can affect the regulation of gene expression .The aim of this study was to predict insulin like growth factor 2 receptor (IGF2R) 3′UTR variants and to test their effects on IGF2R gene expression by bioinformatic analysis . Methods Single nucleotide polymorphisms (SNPs) with minor allele frequency (MAF) in the IGF2R gene 3′UTR were obtained from online databases .The frequency distribution of all the selected IGF2R 3′UTR variants genotypes among the different populations and the linkage disequilibrium ( LD) values of all SNPs were calculated .Additionally , the potential miRNA binding sites were also predicted with the help of online bioinformatic tool . Finally, correlation analysis of the mRNA expression of IGF 2R genotype and different variant genotypes in the lymphoblastoid cell lines was performed. Results In total, 33 SNPs were reported in the 3′UTR, of which only five SNPs (rs8191959, rs200237825, rs3832385, rs201568808, rs1050015) had available minor allele frequency (MAF) values ( >0.05).And only the effect of rs1050015 variant on IGF2R mRNA expression level had significant difference (P=0.010). Conclusion The expression of IGF2R gene can be up-regulated by rs10500105 variant in the 3′UTR, which might support its use as markers of cancer risk and individualized treatment.