Objective:To compare the efficacy of in situ repair and conversion to full-thickness repair in patients with partial tears of the supraspinatus tendon bursa side in rotator cuff tears. Methods:A retrospective analysis was performed on 81 patients who underwent shoulder arthroscopic surgery due to Ellman grade III partial tears on the rotator cuff bursa side in Beijing Friendship Hospital, Capital Medical University from January 2021 to December 2022, according to the different intraoperative supraspinatus tendon repair methods, the patients were divided into the in situ repair group ( n=44) and the partial-to-full-thickness repair group ( n=37). Patients in the in situ repair group were treated with in situ repair for supraspinatus tendon repair, while those in the partial-to-full-thickness repair group were treated with partial-to-full-thickness repair for supraspinatus tendon repair. The general information, pain visual analogue scale (VAS) score, University of California, Los Angeles (UCLA) shoulder joint score and Constant score of the patients were compared and analyzed; the operation time, number of anchors used, and rotator cuff re-tear rate 1 year after surgery were compared and analyzed. The measurement data were expressed as mean ± standard deviation ( ± s), and comparisons between groups were performed using the independent samples t-test. The count data were expressed as the number of cases and percentages, and comparisons between groups were performed using the Chi-square test. Results:All 81 patients completed the follow-up. One year after surgery, the pain VAS scores of the in situ repair group and the partial-to-full-thickness repair group were 1.48±1.07 and 1.38±0.83, respectively, with no significant statistical difference ( P=0.647). The UCLA shoulder joint score and Constant score in the in situ repair group were 30.09±1.46 and 83.05±10.94, respectively, and those in the partial-to-full-thickness repair group were 30.46±1.04 and 84.95±9.20, respectively, there were no significant statistical difference ( P=0.203, 0.405). There was no significant statistical difference in the operation time between the in situ repair group and the partial-to-full-thickness repair group ( P=0.276), but the partial-to-full-thickness repair group was about 11.5 min slower on average. The number of anchors used in the in situ repair group (1.86±0.88) was significantly less than that in the partial-to-full-thickness repair group (2.51±0.65), and the difference was statistically significant ( P<0.001). There was no significant statistical difference in the re-tear rate between the two groups 1 year after surgery ( P=0.625). Conclusions:For patients with partial tears of the supraspinatus tendon bursa side in rotator cuff tears, both in situ repair and partial-to-full-thickness repair can achieve good clinical results, but conversion to full-thickness repair requires longer operation time and more anchors. The choice of specific surgical method needs to be determined based on the patient′s condition and the doctor′s technical proficiency.

This study was aimed at preliminarily investigating the molecular biological functions of the PPE65 protein from Myco-bacterium tuberculosis,and providing foundational data for tuberculosis prevention and control.The basic biological properties of the PPE65 gene-encoded protein were predicted with bioinformatics tools.Sequence information for the Mycobacterium tuberculosis Rv3621c gene and PPE65 protein was retrieved from the NCBI database.The Rv3621c gene was amplified through PCR with the H37Rv genome as a template,then cloned into the pET22b(+)expression vector.The recombinant pET22b(+)-PPE65 plasmid was transformed into Escherichia coli BL21(DE3)competent cells for IPTG-induced expression.Solubility analysis,purification,and identification of the recombinant PPE65 protein were performed.BEAS-2B cells were treated with various concentrations of PPE65 protein for 24 h,and cell proliferation was assessed with CCK-8 assays.PPE65 was found to be composed of 413 amino acids and to have a molecular formula of C????H????N???O???S??,a relative molecular mass of 40 679.88,a theoretical isoelectric point of 4.60,an ali-phatic index of 81.94,and an average hydrophilicity value of 0.319,thus indicating a stable hydrophobic protein lacking signal pep-tides or transmembrane domains.Secondary structure analysis revealed 53.03%α-helix(Hh),2.66%β-sheet(Ee),and 44.31%ran-dom coil(Cc).Bioinformatics predictions identified 38 B-cell epitopes and 22 CTL/Th-cell epitopes.The full-length PPE65 gene(1 308 bp)was confirmed through double restriction enzyme digestion and sequencing,thereby validating the correct construction of the pET22b(+)-PPE65 recombinant plasmid.SDS-PAGE analysis demonstrated that the recombinant protein was found in inclusion bodies,and a single band at 43.7 kDa was observed after purification.Western blotting revealed specific binding to mouse-derived His monoclonal antibodies,thereby confirming successful expression of the PPE65 protein.BEAS-2B cells treated with a PPE65 protein concentration gradient(2.5-20 μg/mL)exhibited a dose-dependent increase in cell number.Compared with those in the PBS control group,TGF-β relative expression levels were significantly higher in all treatment groups(t2.5=4.893,P<0.001,t5.0=4.640,P<0.05,t10=7.535,P<0.05,t20=16.44,P<0.000 1).This study elucidated the structural characteristics of the PPE65 protein,successfully obtained the recombinant protein through prokaryotic expression and purification,and demonstrated its ability to promote BEAS-2B cell proliferation.The underlying mechanism might involve suppression of TGF-β/S mad signaling pathway activation.These findings provide a theoretical basis for understanding the role and regulatory mechanisms of PPE65 during M.tuberculosis infection.

Objective:To characterize the minimal inhibitory concentration (MIC) of macrolides against clinical Mycoplasma pneumoniae (MP) isolates and to investigate the significance of 23S rRNA mutations. Methods:Cross-sectional study.A total of 288 clinical MP strains preserved in the laboratory from 2016 to 2021 were taken for macrolide resistance gene testing and the evaluation of in vitro susceptibility to Azithromycin and Erythromycin.MIC 50 and MIC 90 values were calculated separately for macrolide-susceptible and -resistant strains. Results:All 288 MP strains underwent the test of in vitro susceptibility to Azithromycin, while 86 of them were additionally tested for Erythromycin.Among these strains, 22 strains were Azithromycin-sensitive, and 266 strains were Azithromycin-resistant.A2063G mutations were detected in 260 (97.7%) strains, while A2064G mutations were detected in 6 (2.3%) strains.Azithromycin-resistant strains had an MIC 50 of 128.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 16.000 and 512.000 μg/mL.Seven strains were sensitive and 79 strains were resistant to Erythromycin.Among Erythromycin-resistant strains, A2063G mutations were detected in 73 (92.4%) strains, while A2064G mutations were detected in 6 (7.6%) strains.Erythromycin-resistant strains had an MIC 50 of 256.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 64.000 and 1 024.000 μg/mL. Conclusions:A2063G and A2064G mutations in the 23S rRNA gene of MP are associated with high-level in vitro resistance to Azithromycin and Erythromycin, significantly limiting the clinical effectiveness of these antibiotics.Early resistance gene testing is recommended for suspected MP patients, which can help optimize the treatment, improve prognosis, and prevent resistance spread.

This study was aimed at preliminarily investigating the molecular biological functions of the PPE65 protein from Myco-bacterium tuberculosis,and providing foundational data for tuberculosis prevention and control.The basic biological properties of the PPE65 gene-encoded protein were predicted with bioinformatics tools.Sequence information for the Mycobacterium tuberculosis Rv3621c gene and PPE65 protein was retrieved from the NCBI database.The Rv3621c gene was amplified through PCR with the H37Rv genome as a template,then cloned into the pET22b(+)expression vector.The recombinant pET22b(+)-PPE65 plasmid was transformed into Escherichia coli BL21(DE3)competent cells for IPTG-induced expression.Solubility analysis,purification,and identification of the recombinant PPE65 protein were performed.BEAS-2B cells were treated with various concentrations of PPE65 protein for 24 h,and cell proliferation was assessed with CCK-8 assays.PPE65 was found to be composed of 413 amino acids and to have a molecular formula of C????H????N???O???S??,a relative molecular mass of 40 679.88,a theoretical isoelectric point of 4.60,an ali-phatic index of 81.94,and an average hydrophilicity value of 0.319,thus indicating a stable hydrophobic protein lacking signal pep-tides or transmembrane domains.Secondary structure analysis revealed 53.03%α-helix(Hh),2.66%β-sheet(Ee),and 44.31%ran-dom coil(Cc).Bioinformatics predictions identified 38 B-cell epitopes and 22 CTL/Th-cell epitopes.The full-length PPE65 gene(1 308 bp)was confirmed through double restriction enzyme digestion and sequencing,thereby validating the correct construction of the pET22b(+)-PPE65 recombinant plasmid.SDS-PAGE analysis demonstrated that the recombinant protein was found in inclusion bodies,and a single band at 43.7 kDa was observed after purification.Western blotting revealed specific binding to mouse-derived His monoclonal antibodies,thereby confirming successful expression of the PPE65 protein.BEAS-2B cells treated with a PPE65 protein concentration gradient(2.5-20 μg/mL)exhibited a dose-dependent increase in cell number.Compared with those in the PBS control group,TGF-β relative expression levels were significantly higher in all treatment groups(t2.5=4.893,P<0.001,t5.0=4.640,P<0.05,t10=7.535,P<0.05,t20=16.44,P<0.000 1).This study elucidated the structural characteristics of the PPE65 protein,successfully obtained the recombinant protein through prokaryotic expression and purification,and demonstrated its ability to promote BEAS-2B cell proliferation.The underlying mechanism might involve suppression of TGF-β/S mad signaling pathway activation.These findings provide a theoretical basis for understanding the role and regulatory mechanisms of PPE65 during M.tuberculosis infection.

Objective:To characterize the minimal inhibitory concentration (MIC) of macrolides against clinical Mycoplasma pneumoniae (MP) isolates and to investigate the significance of 23S rRNA mutations. Methods:Cross-sectional study.A total of 288 clinical MP strains preserved in the laboratory from 2016 to 2021 were taken for macrolide resistance gene testing and the evaluation of in vitro susceptibility to Azithromycin and Erythromycin.MIC 50 and MIC 90 values were calculated separately for macrolide-susceptible and -resistant strains. Results:All 288 MP strains underwent the test of in vitro susceptibility to Azithromycin, while 86 of them were additionally tested for Erythromycin.Among these strains, 22 strains were Azithromycin-sensitive, and 266 strains were Azithromycin-resistant.A2063G mutations were detected in 260 (97.7%) strains, while A2064G mutations were detected in 6 (2.3%) strains.Azithromycin-resistant strains had an MIC 50 of 128.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 16.000 and 512.000 μg/mL.Seven strains were sensitive and 79 strains were resistant to Erythromycin.Among Erythromycin-resistant strains, A2063G mutations were detected in 73 (92.4%) strains, while A2064G mutations were detected in 6 (7.6%) strains.Erythromycin-resistant strains had an MIC 50 of 256.000 μg/mL and an MIC 90 of 512.000 μg/mL, with the MIC ranging between 64.000 and 1 024.000 μg/mL. Conclusions:A2063G and A2064G mutations in the 23S rRNA gene of MP are associated with high-level in vitro resistance to Azithromycin and Erythromycin, significantly limiting the clinical effectiveness of these antibiotics.Early resistance gene testing is recommended for suspected MP patients, which can help optimize the treatment, improve prognosis, and prevent resistance spread.

OBJECTIVE：To study spectrum-effect relationship of 11 different solvent extracts from Trollius chinensis against hypoxia/reoxygenation injury of cardiomyocyte . METHODS ：HPLC-MS/MS method was used to establish the fingerprints of 11 different solvent extracts from T. chinensis ，the compounds corresponding to the common peaks were identified by comparing with the substance control and literature information. MTT assay was used to detect the effects of 11 different solvent extracts from T. chinensis on the survival rate of rat myocardial H 9c2 cells injured by hypoxia/reoxygenation. The MDA content ，ROS level in cells and LDH content in the supernatant were detected by ELISA. GRA and PLS method were used to analyze the spectrum-effect relationship between the compounds corresponding to common peak and anti-hypoxia/reoxygenation injury of cardiomyocyte （drug effect）. RESULTS ：There were 22 common peaks in 11 different solvent extracts from T. chinensis ，and 22 compounds were identified. Compared with hypoxia/reoxygenation injury group ，survival rate of hypoxia/reoxygenation injury+S 1-S6，S9 and S 10 groups were increased significantly ，while MDA content ，ROS level and LDH content were decreased significantly （P＜0.05）； ROS level and LDH content of hypoxia/reoxygenation injury+S 8 group w ere decreased significantly （P＜0.05）. The r of GRA analysis of 22 compounds with drug effects were all higher than 0.8. Except for peaks 1，2，7，13，14 and 21，r of PLS analysis of rest peaks with drug effects were higher than 0 发。电话：0431-86058683。E-mial：nml2000@163.com （being positive correlation ）. Top 9 common peaks in the list of contribution rate were peak 6＞11＞4＞5＞8＞9＞12＞10＞15. CONCLUSIONS ：Orientin（peak 6），vitexin（peak 11）， orientin-2″-O-β-L-galacto- pyranosl （peak 4），orientin-2″-O-β-D-Pyrine xylosides （peak 5），quercetin-3-O-glucopyranoside（peak 8），vitexin-2″-O-β-L-galactoside（peak 9），hyperoside（peak 12），vitexin-2″-O-β-D-pyrine xylosides （peak 10），2″-O-（2″′- methylbutyry-loxy）-orientin（peak 15）may be the main components of anti-hypoxia/reoxygenation injury of cardiomyocytes.

Objective:To evaluate the feasibility of transforaminal endoscopic nerve root decompression for degenerative lumbar spinal stenosis (DLSS).Methods: From July 2011 to April 2016,96 cases of single segment DLSS were involved.All the patients had unilateral lower extremity neurological symptoms,signs,neurogenic intermittent claudication of less than 500 m.Imaging examinations (CT or MRI) or diagnostic nerve root block confirmed single segment degeneration.The mean age was (71.6±5.4) years,male: 55 cases,female: 41 cases.Their intraoperative blood loss,operation time,complications,ambulation time and discharge time were recorded.Leg pain VAS,ODI were used to evaluate the pain and lumbar function of the patients.The clinical efficacy was evaluated by Nakai evaluation.Results: All the patients were performed endoscopic decompression of the lateral recess and nerve root by removing the ventral part of the superior facet joint,the ligamentum flavum and the intervertebral disc.The decompression range was from the inferior edge of the upper pedicle to the superior edge of the lower pedicle.The nerve root was detected to have no compression and the pulse of nerve root returned to normal.The patient got ambulant on the operation day and discharged if he had no discomfort symptom.In the study,68 cases got follow up.The mean follow-up time was 12.1 months (6-63 months).The VAS at dif-ferent follow-up time points was improved relative to the baseline,and the difference was statistically significant (F=491.60,P<0.001).The ODI at different follow-up time points was improved relative to the baseline,and the difference was statistically significant (F=189.91,P<0.001).The excellent and good rates of Nakai evaluation were 79.4％ (excellent in 42 cases,good in 12 cases,fair in 10 cases and poor in 4 cases).The mean intraoperative blood loss was (49.29±11.86) mL.The mean operation time was (92.46±21.34) min.The mean ambulation time was 1.8 h.The mean discharge time was 2.3 days.Postoperative epidural hematoma was found in 1 case.Foot drop was found in 1 case.Second stage open surgery was performed in 6 cases.Conclusion: We can apply transforaminal endoscopic decompression for the patients of lumbar spinal stenosis who have unilateral nerve root irritation.Patients with transforaminal endoscopic decompression can get less surgical trauma,quick recovery and obtain good short-term outcome.

Objective To quantifiably measure the methylation frequency of 18 CpG sites in the 3′region of L1 gene and long control region(LCR) gene of HPVl6 DNA,and study the relationship between HPVl6 DNA methylation and severity of cervical lesions. Methods A total of 10 cases Normal/low?grade squamous intraepithelial lesion(Normal/LSIL),10 cases of high?grade squamous intraepithelial lesion(HSIL),and 10 cases of cervical cancer(CC)were recruited for the study. The relationship between severity of cervical lesions and HPV16 DNA methylation was analyzed by bisultlte?pyrosequencing. Results The methylation rate was highest in Normal/LSIL at position 7 089 located in 3′?L1,followed by CC. The low?est was found in HSIL. The difference in methylation percentage among the three lesions was significant(P=0.006). In 7 134,the proportion meth?ylation was also different among three groups(P=0.01),difference in methylation percentage between Normal/LSIL and CC,as well as Normal/LSIL and HSIL was significant(P=0.038,0.017). Conclusion The methylation status of CpG sites 7 089 and 7 134 in the 3′region of L1gene is asso?ciated with the severity of cervical disease. The quantification of HPV DNA methylation can be used for cervical disease screening in clinical samples.

Objective To study the relationship and clinicopathological significance of Numb and epithelial?mesenchymal transition related proteins in human pancreatic cancer( PC)? Methods Sixty?three cases of pancreatic cancer tissues were obtained from department of gastrointestinal surgery in the First Hospital of China Medical University from January 2005 to December 2012, all samples were histopathologically proved to be adenocarcinoma? The expressions of Numb, E?cadherin and Vimentin proteins in 63 cases of pancreatic cancer specimens were detected by immunohistochemistry? Western blot and real?time PCR were used to examine the protein and mRNA levels in two pancreatic cancer cell lines?Pearson and chi?squared tests were used to analyze the relationship and clinicopathological characters with PC patients? Kaplan?Meier curve and log rank test were used to estimate the difference of PC patients′survival? Results The positive rates of Numb,E?cadherin and Vimentin expressions were 46?0%,41?3%and 28?6%, respectively? Numb expression was negatively associated with tumor size, differentiation and UICC stage( r=-0?310, P= 0?010;r=-0?359, P= 0?004;r=-0?228, P=0?020) , while E?cadherin expression was negatively related with tumor differentiation( r=-0?316,P=0?012)? In contrast,Vimentin expression was positively related with pancreatic cancer differentiation and lymph metastasis( r=0?264,P=0?036;r=0?274, P=0?030 )?Correlation analysis showed Numb had a positive association with E?cad expression( r= 0?325, P= 0?010 ) , but had no association with Vimentin? Moreover, patients with co?expression of Numb and E?cadherin had a significantly better overall survival in Kaplan?Meier univariate analysis(P=0?046)?Immunoblotting and real?time PCR showed that high Numb protein and mRNA levels in BxPC?3 cells were followed with high E?cadherin and low Vimentin expressions,whereas low Numb protein and mRNA levels in PANC?1 cells were followed with low E?cadherin and high Vimentin expressions, respectively? Conclusions Numb has a positive relationship with E?cadherin in both pancreatic cancer tissues and cells?The interaction between them might participate in the initiation and development of epithelial?mesenchymal transition in pancreatic cancer.