ObjectiveTo investigate the mechanism through which miR‑21 improves chronic renal fibrosis in mice via targeted modulation of the phosphatase and tensin homolog （PTEN）/protein kinase B （AKT）/mammalian target of rapamycin （mTOR） pathway. MethodsThirty‑two chronic kidney disease model mice were randomly divided into four groups （n=8 each group）： model group， miR‑21 overexpression group， miR‑21 inhibition group， and miR‑21 inhibition + MK‑2206 group. Eight healthy mice were included as the control group. The miR‑21 overexpression， miR‑21 inhibition， and miR‑21 inhibition + MK‑2206 groups received tail‑vein injections of lentivirus （50 μL， 1×10⁸ TU per mouse） once weekly for three weeks. The control and model groups were injected with an equal volume of empty vector （LV‑NC）. The miR‑21 inhibition + MK‑2206 group additionally received gavage of the AKT/mTOR pathway inhibitor MK‑2206 （480 mg/kg） once weekly for three weeks. The expressions of miR‑21， 24 h urinary protein， serum creatinine （Scr）， blood urea nitrogen （BUN）， and renal tissue levels of collagen Ⅰ， collagen Ⅲ， α‑smooth muscle actin （α‑SMA）， and PTEN protein， as well as p‑AKT/AKT and p‑mTOR/mTOR ratios， were compared among groups. HE staining was used to observe pathological changes in renal tissue， and Masson staining was used to observe the degree of renal fibrosis. A dual‑luciferase assay was performed to verify the targeting relationship between miR‑21 and PTEN. ResultsCompared with the model group， miR‑21 expression in renal tissue increased in the miR‑21 overexpression group （P<0.05） and decreased in the miR‑21 inhibition group （P<0.05）. Compared with the model group， the miR‑21 overexpression group showed increased 24 h urinary protein， Scr， BUN， and renal tissue expression of collagen Ⅰ， collagen Ⅲ， and α‑SMA （all P<0.05）， while these indicators decreased in the miR‑21 inhibition group （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group exhibited lower 24‑h urinary protein， Scr， BUN， and renal tissue expression of Collagen Ⅰ， Collagen Ⅲ， and α‑SMA （all P<0.05）. Compared with the model group， the miR‑21 overexpression group showed decreased PTEN protein expression （P<0.05） and increased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， while the miR‑21 inhibition group showed increased PTEN expression （P<0.05） and decreased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group had lower p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， with no significant difference in PTEN protein expression. HE and Masson staining showed normal kidney structure and almost no fibrosis in the control group. The model group exhibited glomerular enlargement， capillary loop adhesion， and focal fibrosis. The miR-21 overexpression group showed severe destruction of glomerular structure， accompanied by extensive fibrosis and renal tubular atrophy. The pathological changes and degree of fibrosis were alleviated in the miR-21 inhibition group. The miR-21 inhibition + MK-2206 group showed only mild pathological changes and mild fibrosis， with the interstitium being largely normal. Compared with PTEN-WT + NC mimics 1， the relative luciferase activity in the PTEN-WT + miR-21 mimics group decreased （P<0.001）. There was no statistically significant difference in relative luciferase activity between PTEN-WT + NC mimics group and PTEN-MUT + miR-21 mimics group. ConclusionmiR‑21 may improve renal function indicators and alleviate renal fibrosis in chronic kidney disease mice via targeted modulation of PTEN and subsequently inhibiting the AKT/mTOR pathway.

To assess the cross-contamination risk and genetic evolution ofStaphylococcus aureus(S.aureus)at various stages in a swine slaughterhouse,and to provide a reference for risk evalua-tion and control measures of S.aureus contamination during swine slaughtering,we conducted whole-genome sequencing on 31 isolates of S.aureus collected and preserved from the slaughter-house.Bioinformatics analyses were performed to investigate the genomic characteristics and genet-ic relationships of these isolates.Results revealed cross-contamination across different slaughter-house stages,predominantly with ST398-t1451 strains.Additionally,highly virulent ST9-t899 strains and ST398-t11 strains with numerous resistance genes were detected at various stages.The strains predominantly carried virulence genes such as hlgA,hlgB,and hlgC,with varying num-bers of resistance genes.Notably,two strains carried the optrA gene and three strains carried the cfr gene;the presence of the optrA gene is relatively rare among S.aureus.These genes confer re-sistance to novel synthetic antibiotics such as oxazolidinones and florfenicol and have the potential for horizontal gene transfer,increasing the risk of dissemination both within and beyond the slaughterhouse.Importantly,the study also detected a LA-MRSA-ST9 strain in water samples from the slaughterhouse,which could potentially infect humans.This strain exhibits zoonotic char-acteristics,highlighting the need for stricter protective measures for slaughterhouse workers to mitigate occupational exposure and infection risks.

Objective Piperazine derivatives are a group of emerging psychoactive substances with excitatory and hallucinogenic effects on the central nervous system.This study established a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)screening method for the detection of 40 piperazine compounds in urine.Methods A 200 μL urine sample(spiked with an internal standard at 1 ng/mL)was subjected to liquid-liquid extraction with ethyl acetate.After nitrogen evaporation,the residue was redissolved in 200 μL methanol and injected for analysis.Separation was performed on a Waters Acquity UPLC? HSS T3 column(100 mm × 2.1 mm,1.8 μm).The mobile phase consisted of(A)20 mmol/L ammonium acetate buffer containing 0.1％formic acid and 5％acetonitrile,and(B)acetonitrile.Gradient elution was applied,and detection was carried out in multiple reaction monitoring(MRM)mode.Quantification was achieved using an internal standard calibration curve.Results The 40 piperazine substances demonstrated good linearity within the range of 1-50 ng/mL,with correlation coefficients of 0.995-0.998.The extraction recovery ranged from 51.51％to 104.1％.Intra-day precision was below 5％,while inter-day precision ranged from 1.61％to 10.17％.Accuracy was between-7.84％and 8.77％.The limits of detection were 0.2-1 ng/mL,and the limit of quantification was 1 ng/mL.Conclusion The proposed method requires only a small sample volume,exhibits high sensitivity,selectivity,and stability,and offers short run times.It is suitable for the qualitative and quantitative determination of piperazine derivatives in urine in forensic toxicology practice.

Objective:To evaluate the feasibility and safety of endoscopic retrograde cholangiopancreatography (ERCP) combined with peroral cholangioscopy in the treatment of gallbladder stones with common bile duct stones.Methods:Clinical data of 19 patients with gallbladder stones and common bile duct stones admitted to the Digestive Endoscopy Center, Jilin People’s Hospital from July 2019 to November 2024 were retrospectively analyzed, including 13 males and 6 females, aged (68.2±14.2) years. All patients underwent ERCP combined with peroral cholangioscopy. Perioperative data, including the long diameter of the common bile duct stone, the long diameter of the gallbladder stone, the number of stones, ERCP operative time, gallbladder stone extraction time, stone clearance status, hospitalization duration, and complications, were recorded. Postoperative follow-up was conducted through outpatient visits or telephone consultations to monitor recurrence.Results:The long diameter of the common bile duct stones was (9.55±2.86) mm, and the long diameter of the gallbladder stones was 4.0(3.0, 5.5) mm. Among the 19 patients, 5 had single gallbladder stones, and 14 had multiple stones. The ERCP operative time was (49.0±18.4) min, and the gallbladder stone extraction time was (25.0±11.7) min. The methods used for handling the gallbladder neck were as follows: two cases involved stone removal after 6 mm balloon dilation; six cases involved stone removal after metal stent placement; three cases involved the placement of a plastic stent in the gallbladder for three months, followed by stone extraction using ERCP combined with peroral cholangioscopy; eight cases were treated directly for stone removal without specific procedures. Among the 19 patients, 11 completed stone removal within one treatment, while eight required a second treatment. All patients had complete clearance of both common bile duct and gallbladder stones under direct peroral cholangioscopy. No severe complications occurred postoperatively, although two cases developed post-ERCP pancreatitis. The postoperative hospitalization time was 8.0 (6.0, 10.0) d. The follow-up duration for the 19 patients was 14.0 (8.5, 20.0) months. One patient had a recurrence of gallbladder stones 12 months postoperatively, while no other patients had recurrence at the final follow-up.Conclusion:ERCP combined with peroral cholangioscopy is a safe and feasible approach for treating gallbladder stones with common bile duct stones.

Objective:To evaluate the feasibility and safety of endoscopic retrograde cholangiopancreatography (ERCP) combined with peroral cholangioscopy in the treatment of gallbladder stones with common bile duct stones.Methods:Clinical data of 19 patients with gallbladder stones and common bile duct stones admitted to the Digestive Endoscopy Center, Jilin People’s Hospital from July 2019 to November 2024 were retrospectively analyzed, including 13 males and 6 females, aged (68.2±14.2) years. All patients underwent ERCP combined with peroral cholangioscopy. Perioperative data, including the long diameter of the common bile duct stone, the long diameter of the gallbladder stone, the number of stones, ERCP operative time, gallbladder stone extraction time, stone clearance status, hospitalization duration, and complications, were recorded. Postoperative follow-up was conducted through outpatient visits or telephone consultations to monitor recurrence.Results:The long diameter of the common bile duct stones was (9.55±2.86) mm, and the long diameter of the gallbladder stones was 4.0(3.0, 5.5) mm. Among the 19 patients, 5 had single gallbladder stones, and 14 had multiple stones. The ERCP operative time was (49.0±18.4) min, and the gallbladder stone extraction time was (25.0±11.7) min. The methods used for handling the gallbladder neck were as follows: two cases involved stone removal after 6 mm balloon dilation; six cases involved stone removal after metal stent placement; three cases involved the placement of a plastic stent in the gallbladder for three months, followed by stone extraction using ERCP combined with peroral cholangioscopy; eight cases were treated directly for stone removal without specific procedures. Among the 19 patients, 11 completed stone removal within one treatment, while eight required a second treatment. All patients had complete clearance of both common bile duct and gallbladder stones under direct peroral cholangioscopy. No severe complications occurred postoperatively, although two cases developed post-ERCP pancreatitis. The postoperative hospitalization time was 8.0 (6.0, 10.0) d. The follow-up duration for the 19 patients was 14.0 (8.5, 20.0) months. One patient had a recurrence of gallbladder stones 12 months postoperatively, while no other patients had recurrence at the final follow-up.Conclusion:ERCP combined with peroral cholangioscopy is a safe and feasible approach for treating gallbladder stones with common bile duct stones.

Objective:To investigate the expression characteristics of Zeste enhancer of Zeste homolog 2 (EZH2) in breast cancer tissue and its influence on tumor progression and prognosis.Methods:Transcriptome data of breast cancer tissue and normal breast tissue adjacent to cancer as well as clinical data of patients were obtained from the cancer genome atlas (TCGA) database, gene expression comprehensive database and European genome phenotype archives database, and the difference of EZH2 expression was analyzed using TIMER 2.0 platform. The survival information of breast cancer patients was obtained from the Kaplan Meier Plotter database, and the overall survival time, relapse free survival time and distant metastasis free survival time of breast cancer patients with low EZH2 expression and high EZH2 expression were compared. Select 14 nude mice were selected and randomly divided into si-EZH2 group and control group, with 7 mice in each group.MCF7 culture suspensions transfected with EZH2 knockdown plasmid and control plasmid were inoculated for corresponding group. The body mass and tumor volume of two groups of nude mice inoculated with MCF7 cells were compared at different times. On the 28th day, the nude mice were euthanized and the tumors were dissected to compare the tumor mass of the two groups of nude mice. The normally distributed quantitative data was represented by xˉ ± s. Two independent sample t-tests were used for comparison between two groups, repeated measures ANOVA was used for comparison of body mass and tumor volume between two groups of nude mice at different times, and Bonferroni test was used for pairwise comparison. The comparison of survival rates was conducted using log rank test. Results:A total of 1085 breast cancer tissues and 291 normal adjacent breast tissues were included in the TCGA database. EZH2 expression in breast cancer tissues was higher than that in normal adjacent breast tissues ( P<0.05). In the Kaplan Meier Plotter database, the total survival time, relapse free survival time, and distant metastasis free survival time of breast cancer patients in the EZH2 overexpression group were shorter than those in the EZH2 low expression group ( P=0.013, <0.001, <0.001). After 7 days of inoculation with MCF7 culture suspension, significant subcutaneous tumors were observed on the left back of both groups of nude mice. On the first day, there were no statistically significant difference in body mass between the two groups of nude mice ( P>0.05); On day 7, 13, 19, 25, and 28, the body mass and tumor volume of both groups of nude mice gradually increased (nude mouse body mass: within group F=29.31, P<0.001, between groups F=234.32, P<0.001, Finteraction=16.83, P<0.001; Tumor volume: within group F=34.00, P<0.001, between groups F=193.17, P<0.001, Finteraction=35.61, P<0.001). And the body mass of the siEZH2 group nude mice was higher than that of control group (all P<0.05). On days 19, 25, and 28, tumor the volume of the siEZH2 group nude mice was smaller than that of control group (all P<0.05). On the 28th day, the mass of tumors dissected in the siEZH2 group of nude mice was lower than that in the control group [(0.30±0.07) g vs. (0.61±0.14) g, t=5.16, P<0.001]。 Conclusions:EZH2 is highly expressed in breast cancer tissues and is significantly associated with poor prognosis. Knockdown of EZH2 can significantly inhibit the proliferation and tumor formation of breast cancer cells.

Objective:To elucidate the genomic characteristics of the immunoglobulin (IG) heavy-chain variable region and light-chain variable region, the expression of subclones, and the prognostic significance in patients with CLL.Methods:Blood and/or bone marrow specimens were gathered from a cohort of 36 patients with CLL diagnosed at Jiangsu Province Hospital from December 2018 to May 2023, including 12 cases of B cell receptor (BCR) stereotyped patients. IG heavy-chain (IGH) and light-chain (IG Kappa [IGK] and IG lambda [IGL]) gene rearrangements were performed using next-generation sequencing (NGS) technology to analyze the characteristics and prognostic value in CLL.Results:NGS detection of IG variable region (IGHV) demonstrated a significant correlation and superior consistency with Sanger sequencing ( r=0.957, P < 0.001). Among the 36 patients, the IGH variant (IGHV) was observed in 9 (25.0%) but not in 27 (75.0%) participants. The incidence of the MYD88 mutation was higher among patients with mutated IGHV [1/27 (3.7%) vs 4/9 (44.4%), P=0.00]. A high incidence of trisomy 12 was observed in the IGHV #8/#8B subset [4/11 (36.4%) vs 1/25 (4.0%), P=0.023], which were more likely to develop Richter transformation [8/11 (72.7%) vs 4/25 (16.0%), P=0.002]. In the patient cohort, 36 individuals (36/36, 100.0%) used the IGK variable, whereas 15 individuals (15/36, 41.7%) employed the IGL variable (IGLV). IGLV3 - 21 reported the highest utilization rate in IGLV (5/15, 33.3%). Remarkably, patients with CLL with IGLV3-21 fragments were exclusively observed in the Binet C stage and Rai Phase Ⅲ-Ⅳ, with an incidence of del (13) (q14) at 60.0% (3/5). The median time to first treatment (TTFT) of patients with or without IGLV3 - 21 fragments was 5.2 (1.1 - 41.5) and 9.9 (0.1 - 94.4) months, respectively. Using the total reads threshold of 2.5%, 4 (4/36, 11.1%) samples were detected to have two IGHV productive clones. The median TTFT and overall survival (OS) time were 2.8 (0.9-72.7) and 12.8 months in patients with one mutated clone and 57.5 (32.0-120.7) and 51.8 months in those with two mutated clones, respectively. The median TTFT and OS time were 10.9 (0.3-94.4) and 6.3 (0.1 - 12.5) months in patients with one unmutated clone and 49.9 (22.2 - 211.1) and 30.0 (9.6 - 50.3) months in those with multiple unmutated clones, respectively ( P>0.05) . Conclusions:Detection of IG gene rearrangements using NGS technology not only facilitates the analysis of the IGHV mutation status, dominant clones, and prognostic value but also contributes to the exploration of IGK/IGL gene rearrangement fragments and the utilization of subclones. Further, it provides information about the poor prognosis of IGLV3 - 21 CLL. The shortened survival of the two unmutated clone groups in the IGHV unmutated group may indicate a poor prognosis.

Objective To explore the optimisation path of medical resource allocation efficiency improvement in Traditional Chinese Medicine (TCM) hospitals under the synergistic effect of multiple factors,so as to provide powerful support for the balanced development of medical resources in TCM hospitals.Methods The comprehensive efficiency of resource allocation in TCM hospitals in 31 provinces (cities and districts) of China obtained by Data Envelopment Analysis was taken as the outcome variable,and with the help of Fuzzy-set Qualitative Comparative Analysis,it examined the configurations of efficient medical resource allocation,considering the following conditional variables:per capital Regional GDP,the proportion of fiscal allocation revenue to total income,the ratio of TCM practicing (assistant) physicians to all practicing (assistant) physicians,the average length of hospital stay for discharged patients,the number of total diagnosis and treatments per thousand population,and the number of hospital beds per thousand population.Results By analyzing the conditional patterns of efficient allocation of medical resources in TCM hospitals,three equivalent driving paths can be summarized,which are comprehensive service capacity,TCM advantage and hospitalization driving paths.Conclusion The overall level of medical resource allocation efficiency of TCM hospitals in China needs to be improved.In the future,efforts should be made to improve the comprehensive service capacity and operational efficiency of hospitals,give full play to the advantages of TCM,build a high-quality TCM talent team,reasonably shorten the average hospital stay,and improve the utilization efficiency of hospital beds.

Objective:To study the development status and characteristics of the Traditional Chinese Medicine(TCM)service capacity of primary medical institutions(referred as primary)in Beijing.Methods:The case-base aggregation method was used to analyze the total amount,distribution,and development changes of total TCM costs in primary healthcare institutions in Beijing.Results:From 2019 to 2023,the average annual growth rate of the TCM costs of primary healthcare institutions in Beijing was 10.09％,and the proportion in total TCM costs increased from 16.01％to 18.19％.The average annual growth rate of the TCM costs of community health service centers(stations)was 11.78％,but there were negative growths in outpatient departments,clinics,and village clinics.The proportion of the primary TCM costs in its health costs increased from 29.16％to 32.56％.The average annual growth rate of income from primary herbal medicines and processed Chinese medicines were 16.75％and 10.13％.Conclusion:The primary TCM undertaking has developed steadily,and Beijing should continue to guide high-quality TCM resources to sink.Multiple measures should be taken to promote the development of TCM in outpatient departments,clinics,and village clinics.The coordination between prices and medical insurance reimbursement and payment methods should be strengthened.Supervision over the quality and use of Chinese medicine should be strengthened,and the technical service capabilities of primary TCM should be enhanced.

To assess the cross-contamination risk and genetic evolution ofStaphylococcus aureus(S.aureus)at various stages in a swine slaughterhouse,and to provide a reference for risk evalua-tion and control measures of S.aureus contamination during swine slaughtering,we conducted whole-genome sequencing on 31 isolates of S.aureus collected and preserved from the slaughter-house.Bioinformatics analyses were performed to investigate the genomic characteristics and genet-ic relationships of these isolates.Results revealed cross-contamination across different slaughter-house stages,predominantly with ST398-t1451 strains.Additionally,highly virulent ST9-t899 strains and ST398-t11 strains with numerous resistance genes were detected at various stages.The strains predominantly carried virulence genes such as hlgA,hlgB,and hlgC,with varying num-bers of resistance genes.Notably,two strains carried the optrA gene and three strains carried the cfr gene;the presence of the optrA gene is relatively rare among S.aureus.These genes confer re-sistance to novel synthetic antibiotics such as oxazolidinones and florfenicol and have the potential for horizontal gene transfer,increasing the risk of dissemination both within and beyond the slaughterhouse.Importantly,the study also detected a LA-MRSA-ST9 strain in water samples from the slaughterhouse,which could potentially infect humans.This strain exhibits zoonotic char-acteristics,highlighting the need for stricter protective measures for slaughterhouse workers to mitigate occupational exposure and infection risks.