Objective To explore the action targets and related mechanisms of pogostemon cablin in the treatment of glioma based on network pharmacology and cellular experiments.Methods The active ingredi-ents and action targets of pogostemon cablin were collected by using databases such as the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the Traditional Chinese Med-icine Comprehensive Database(TCMID).The relevant targets of glioma were collected through databases such as Gene Cards and OMIM,and the network diagrams of"drug-target"and"disease-target"were estab-lished by using Cytoscape3.7.1 software.The intersection targets of pogostemon cablin for treating glioma were obtained and STRING database was used to analyzed,and their protein-protein interaction(PPI)net-work diagram was established.Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis on key targets were performed by using the DAVID database.In vitro cell experiments on key signaling pathways were conducted,U87 glioma cells were collected and divided into the control group(added to basic culture medium)and the 60,90,120 μg/mL pog-ostemon cablin groups(added to culture medium containing 60,90,and 120 μg/mL pogostemon cablin,respec-tively).CCK-8 assay and wound healing assay were used to detect the proliferation and migration ability of each group of cells,and the protein expression levels of mitogen activated protein kinase 1(MAPK1),recom-binant V-rel reticuloendotheliosis viral oncogene homolog A(RELA)phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),phosphorylated-PI3K(p-PI3K),and phosphorylated-Akt(p-Akt)in each group of cells were detected by Western blot.Results A total of 94 main chemical constituents of pogostemon cablin were obtained.According to the screening conditions,9 effective active ingredients,239 drug target proteins,4 526 glioma related genes and 121 drug and disease common targets were identified.Protein interaction net-works discovered that Akt1,c-Jun N-terminal kinase(JUN),MAPK1,RELA,interleukin(IL)-6 and epider-mal growth factor receptor(EGFR)may be the key targets of pogostemon cablin in the treatment of glioma.GO function enrichment analysis identified 1 819 items in biological processes,109 items of cellular compo-nents and 216 items of molecular functions,which involved various aspects such as the positive regulation of the cellular metabolic process,the activity of protein homodimers,kinase activation,and membrane rafts.KEGG pathway enrichment analysis identified 181 related signaling pathways,involving anti-cancer,inflamma-tion,cell apoptosis,endocrine,immune regulation and so on,mainly including the PI3K/Akt pathway,ad-vanced glycation advanced glycation end products(AGE)/recepter for advanced glycation end products(RAGE)pathway,IL-6 pathway,and HIF-1 pathway,TNF pathway and so on.The in vitro experimental re-sults showed that compared with the control group,with the increase of the dose of the alcohol extract of pog-ostemon cablin,the cell activity decreased,the cell migration ability decreased,and the expression levels of MAPK1,RELA,p-PI3K and p-Akt proteins decreased(P＜0.05).Conclusion Pogostemon cablin can syner-gistically inhibit the development of glioma through multiple targets and pathways,and can suppress cell pro-liferation of U87 cells through the PI3K/Akt pathway.

OBJECTIVE To evaluate the effictiveness and safety of recombinant human endostatin combined with afatinib and teggio in the treatment of advanced lung squamous cell carcinoma. METHODS A total of 25 patients with driver-negative advanced lung squamous cell carcinoma were included in this single-arm prospective study, and the enrolled patients were treated with recombinant human endostatin combined with afatinib and teggio as scheduled. Progression-free survival(PFS), overall survival(OS), disease control rate(DCR), objective response rate(ORR), and adverse reactions(AR) were observed and analyzed. RESULTS The 25 enrolled patients received at least 2 cycles of second-line treatment, and were followed up as of March 31, 2023. Among them, 4 patients had partial remission, 17 patients had stable disease, and 4 patients experienced progressive disease. The ORR confirmed by the researchers was 16%(95%CI, 4.5%−36.1%), DCR was 84%(95%CI, 63.9%−95.5%), and median PFS was 5.3 months(95%CI, 3.5−6.9 months). The median OS had not yet been achieved. The entire group of patients had good treatment tolerance, and the most common level Ⅲ or Ⅳ adverse events related to treatment were leukopenia(20%) and rash(12%), with no reported treatment-related deaths. CONCLUSION Recombinant human endostatin combined with afatinib and teggio in the second line treatment of advanced lung squamous cell carcinoma can prolong the progression free survival period of patients and is relatively safe, which is worth further exploration and promotion.

Objective To investigate the application value of the MHSeqTyper47 kit in kinship identification.Methods Multiplexed amplification and library preparation were performed for DNA samples from 113 related individuals by using the MHSeqTyper47 kit.The libraries were sequenced on a MiSeq FGx sequencer,and the data were analyzed using MHTyper for microhaplotype genotyping.The kinship indexes were calculated to evaluate the application efficiency of this kit in kinship identification and compared with those of the GlobalFilerTM kit.Results For the MHSeqTyper47 kit,the CPI values in trio identification were 1.43× 1011～6.15×1018.The CPI values in duo identification were 1.02× 105～1.53× 1013.The CFSI values in full sibling identification were 7.73×101～2.59×1016.Trios,duos and full siblings could be completely distinguished from unrelated pairs.The combined efficiency of these two kits in 2nd-degree kinship identification was 0.466 2.Conclusion The application value of MHSeqTyper47 kit is relatively higher in the identification of lst-degree kinships.If jointly used with the GlobalFilerrM kit,2nd-degree kinship identification could be achieved in some cases.

Objective To explore the expression of mocro RNA(miR)-142-5p and suppressor of cytokine signaling 1(SOCS1)mRNA in peripheral blood mononuclear cells(PBMC)of mice and clinical patients with ankylosing spondylitis(AS)and their impact on immune function.Methods The mRNA levels of miR-142-5p and SOCS1 in PBMC of 30 patients with AS(Patient group)and 30 healthy controls(Health group)treated in the First People's Hospital of Shangqiu from January 2022 to March 2023 were measured by quantitative real time PCR(qRT-PCR).AS mice models were induced by bovine proteoglycan combined with complete Freund's adjuvant,and these mice were divided into control group,model group,NC group and antagomir group.Normal saline was injected into tail vein in control group and model group,and NC-antagomir and miR-142-5p-antagomir were injected into tail vein in NC group and antagomir group,respectively.After 2 weeks of treatment,the arthritis symptom scores of mice in each group were evaluated.The morphology of ankle joint was evaluated by hematoxylin eosin(HE)staining.The levels of Th1 cytokine interferon-γ(IFN-γ),Th2 cytokine interleukin-4(IL-4),Th17 cytokine interleukin-17(IL-17)and Treg cytokine forkhead box protein P3(FOXP3)in PBMC of mice were detected by ELISA method.The mRNA and protein expression of miR-142-5p,SOCS1,IFN-γ,IL-4,IL-17 and FOXP3 in PBMC and ankle joints were detected by qRT-PCR and Western blot.Results Compared with Health group,the level of miR-142-5p in PBMC of patient group was increased(1.00±0.21 vs 3.03±0.99,t=10.997,P＜0.001),while the level of SOCS1 mRNA was decreased(1.00±0.18 vs 0.41±0.09,t=15.956,P＜0.001).Compared with control group,miR-142-5p level(1.00±0.04 vs 4.00±0.52)and the mRNA and protein levels of IFN-γ and IL-17 in ankle joint tissue of model group were increased,while the mRNA and protein levels of SOCS1,IL-4 and FOXP3 were decreased,with significant differences(t=23.356,31.420,48.056,47.224,38.035,29.007,54.183,28.123,55.155,26.758,45.346,all P＜0.05).The arthritis symptom score was increased(7.83±0.94 vs 0.00±0.00,t=22.212,P＜0.05),and the ankle joint structure was damaged.Serum IFN-γ,IL-17 levels,IFN-γ/IL-4 ratio(0.81±0.08 vs 2.08±0.33)and IL-17/FOXP3 ratio(0.41±0.03 vs 1.27±0.10)were increased,and the differences were statistically significant(t=15.382,35.779,8.934,35.130,all P＜0.05).Compared with NC group,miR-142-5p level(3.89±0.33 vs 1.47±0.10),the mRNA and protein levels of IFN-γ and IL-17 in ankle joint tissue of antagomir group were decreased,while the mRNA and protein levels of SOCS1,IL-4 and FOXP3 were increased,and the differences were statistically significant(t=18.846,22.969,43.454,32.617,23.259,20.881,41.832,11.994,32.977,15.190,35.834,all P＜0.05).The arthritis symptom score was decreased(7.42±1.24 vs 2.75±0.75,t=13.233,P＜0.05),and the shape of the ankle joint of the rats was improved.Serum IFN-γ,IL-17 levels,IFN-γ/IL-4 ratio(1.22±0.11 vs 1.91±0.19)and IL-17/FOXP3 ratio(0.69±0.05 vs 1.23±0.12)were decreased,and the differences were statistically significant(t=8.688,22.972,3.785,22.007,all P＜0.05).Conclusion MiR-142-5p was highly expressed in AS.Down-regulation of miR-142-5p using antagonists may reduce Th1/Th2 ratio and Th17/Treg ratio through up-regulation of SOCS1,there by improving the immune balance of AS mice and inhibiting the progression of AS.

Massive rotator cuff tears, with mean tendon tears>5 cm or involving two or more tendons in a tear, often result in shoulder pain, stiffness, limited range of motion and impaired function, significantly impacting the patients′ quality of life. At present, massive rotator cuff tears are mainly treated surgically with joint cleaning, rotator cuff repair and patch graft repair, etc, but the problems such as tendon end retraction and high rotator tension after surgical repair lead to high postoperative retear rate. The autogenous long head of biceps tendon (LHBT), as an autograft which has the advantages of high accessibility and low cost, has been employed in the repair of massive rotator cuff tears. However, there is still controversy in the choice of specific surgical method. To this end, the authors reviewed the research progress on the application of LHBT in the repair of massive rotator cuff tears, aiming to provide reference for its clinical treatment.

Objective:To compare the efficacy of superior capsular reconstruction using a "sandwich" patch graft versus a fascia lata autograft in the treatment of irreparable massive rotator cuff tears (IMRCTs).Methods:A retrospective analysis was conducted on 50 patients with IMRCTs who underwent superior capsule reconstruction at Ningbo Medical Center Li Huili Hospital from January 2019 to May 2021. Patients were categorized into two groups based on the type of graft used: the "sandwich" group (27 patients), utilizing a "sandwich" patch graft, and the fascia lata group (23 patients), utilizing a simple fascia lata graft. The "sandwich" group consisted of 10 males and 17 females with a mean age of 65.6±5.7 years (range, 55-76 years), including 6 cases on the left shoulder and 21 cases on the right shoulder. The fascia lata group comprised 10 males and 13 females with a mean age of 65.5±4.2 years (range, 56-72 years), including 4 cases on the left shoulder and 19 cases on the right shoulder. Clinical outcomes were assessed preoperatively and at 1 week, 6 months, 1 year, and 2 years postoperatively using the American Shoulder and Elbow Surgeons (ASES) score, the University of California Los Angeles (UCLA) score, and the visual analogue scale (VAS) for pain and shoulder activity. Imaging outcomes, including acromio humeral distance (AHD) and graft status, were evaluated via radiographs or MRI.Results:The follow-up duration was 40.0±8.4 months (range, 26-54 months) for all patients. The "sandwich" group demonstrated significantly better outcomes compared to the fascia lata group. Improvements were noted in the ASES score (90.1±8.7 vs. 66.8±22.0), ASES score improvement (58.0±11.8 vs. 36.7±24.2), UCLA score (31.0±3.1 vs. 23.0±8.7), UCLA score improvement (20.1±4.5 vs. 12.7±9.2), active elevation (160.0°±21.3° vs. 124.8°±37.4°), active elevation improvement (70.2°±31.4° vs. 33.7°±42.4°), external rotation (35.0°±9.0° vs. 29.0°±6.9°), external rotation improvement (11.3°±7.3° vs. 7.2°±10.4°), AHD (6.4±1.5 mm vs. 4.4±1.0 mm), AHD improvement (3.0±1.6 mm vs. 1.5±1.0 mm), the difference was statistically significant ( P<0.05). The graft healing rate of the "sandwich" group reached 93%, which was higher than the 74% in the fascia lata group, but the difference was not statistically significant (χ 2=1.984, P=0.159). One case of subcutaneous hematoma was reported in the fascia lata group postoperatively. Additionally, two patients in the fascia lata group reported mild thigh discomfort at the last follow-up, which did not impair walking. No cases of infection, joint stiffness, or vascular or nerve damage were observed. Conclusion:Superior capsular reconstruction using a "sandwich" patch graft significantly enhances the healing rate of grafts and short-term postoperative clinical outcomes in patients with irreparable massive rotator cuff tears.

Objective:To explore and analyze the changes of miRNA-138 and miRNA-26b before and after transsphenoidal surgery for pituitary tumors and their clinical significance.Methods:86 patients with functional pituitary tumors who underwent transsphenoidal surgery in Linyi People’s Hospital from Apr. 2020 to Apr. 2021 were retrospectively analyzed. The recurrence within 1 year after operation was followed up and the patients were divided into unrecovered group (24 cases) and cured group (62 cases) . Clinical data such as age, gender, tumor pathological type, Knosp grade, first operation, tumor diameter, intraoperative tumor residue, Ki-67 and adjuvant therapy were collected. Fasting venous blood was collected before surgery and the next morning after surgery. Quantitative real-time PCR was used to detect the serum levels of mircoRNA-138 (miRNA-138) and miRNA-26b (mirNA-26B) , and the changes of serum miRNA-138 and miRNA-26b levels before and after surgery. Univariate and multivariate logistic regression were used to analyze the relationship between serum miRNA-138 and miRNA-26b levels and postoperative prognosis, and ROC curve was drawn to analyze their predictive value for postoperative prognosis.Results:Univariate analysis showed that Knosp grade, tumor diameter, intraoperative tumor residual, Ki-67, and adjuvant therapy were associated with the recurrence of body tumor after transsphenoidal surgery ( P<0.05) . After operation, the expression of miRNA-138 in serum was higher than that before operation, and the expression of miRNA-138 in unhealed group (4.13±1.12) was higher than that in cured group (3.56±0.84) ( P<0.05) . The expression of miRNA-26b in serum after operation was lower than that before operation, and the expression of miRNA-26b in the unhealed group (2.34±0.62) was lower than that in the cured group (2.75±0.58) ( P<0.05) . Pituitary tumor hormone increased before operation and returned to normal after operation. Multivariate logistic regression analysis showed that tumor diameter ≥40cm (OR=3.476, 95%CI: 1.267-9.539) , postoperative tumor residual (OR=3.155, 95%CI: 1.236-8.052) , Ki-67≥3% (OR=3.885, 95%CI: 2.038-7.403) , postoperative serum miRNA-138 expression ≤3.62 (OR=2.323, 95%CI: 1.536-3.513) , postoperative serum miRNA-26b expression ≥2.59 (OR0.453, 95 %CI: 0.286-0.717) was an independent risk factor for the recurrence of pituitary tumors after transsphenoidal surgery ( P<0.05) . When the optimal cut-off value of serum miRNA-138 was 3.62, the area under the curve for predicting the recurrence of pituitary tumors after transsphenoidal surgery was 0.78, the sensitivity was 81.35%, and the specificity was 71.46%; the optimal cut-off value of serum miRNA-26b When it was 2.59, the area under the curve for predicting and predicting the recurrence of pituitary tumors after transsphenoidal surgery was 0.75, at this time, the sensitivity was 78.62%, and the specificity was 72.33%. The lower area was 0.83, the sensitivity was 85.47%, and the specificity was 72.38%. Conclusion:The expression of serum miRNA-138 was up-regulated and the expression of miRNA-26b was down-regulated after transsphenoidal surgery for pituitary tumors, and its abnormal expression was associated with postoperative recurrence, which has a good predictive value for predicting postoperative recurrence.

Objective:To investigate the effect of arthroscopic superior capsular reconstruction (ASCR) of irreparable massive rotator cuff tears (RCTs) using the "Sandwich" patch graft (autologous fascia lata + LARS artificial ligament + autologous fascia lata).Methods:The patients with irreparable massive RCTs who underwent ASCR by using "Sandwich" patch graft were retrospectively evaluated between December 2016 and October 2018. All cases were followed up more than two years. The pain and function of the shoulder were evaluated by visual analogue scale (VAS), active forward elevation (aFE), American Shoulder & Elbow Surgeons score (ASES), University of California Los Angeles (UCLA) score, and Constant-Murley score. The acromiohumeral distance (AHD) and patch healing were assessed by shoulder X-ray and MRI scan.Results:Twenty-nine patients (12 males and 17 females) were enrolled for final analysis. The average age was 66.0±5.88 years (range 55-77 years). The average length of follow-up was 35.3±7.20 months (range 24-46 months). The tendon of the subscapularis muscle was intacted in 13 cases and repairable in 16 cases. All patients' teres minor muscles were intact and the function of deltoid muscles was all complete. Based on Hamada classification of massive RCTs, 11 cases with type 2, 14 cases with type 3, and 4 cases with type 4. At the last follow-up visit, the AFE of the surgical shoulder was 158.45°±23.87° (range 70°-180°), which was significantly higher than before the surgery 92.59°±45.99° (range 20°-160°, t=6.190, P<0.001). The ASES score was 92.92±9.08 (range 64-100), UCLA score was 31.62±3.93 (range 19-35), and Constant-Murley score was 85.8±8.44 (range 68-94), which were higher than those before the operation 30.69±12.99 (range 68-95), 10.93±3.43 (range 4-17), 39.62±12.68 (range 14-55). There were significant differences between them, respectively ( t=21.145, P<0.001; t=21.348, P<0.001; t=16.333, P<0.001). The VAS was 0.41±0.57 (range 0-2 points), which was significantly lower than that of 4.90±1.05 (range 3-7 points) before operation ( t=20.267, P<0.001). At the last follow-up visit, MRI showed that the AHD increased from 3.31±1.57 mm (range 1.1-6.6 mm) to 6.94±1.76 mm (range 3.0-10.8 mm) significantly ( t=12.195, P<0.001). Radiological outcomes were evaluated according to MRI, the total healing rate was 89.7% (26/29). There were two cases of complete tears, which including one case of infection and 1 partial tears. Conclusion:ASCR of irreparable massive RCTs using "Sandwich" patch graft showed the high healing rate in the short-term follow-up, which is possible to restore the shoulder functions early. It is an effective method for the treatment of irreparable massive RCTs.

Objective: To investigate the clinical outcomes of arthroscopic bridging reconstruction of irreparable massive rotator cuff tears using autogenous fascia lata. Methods: From July 2015 to July 2017, a total of 10 cases (4 male and 6 female) who were treated with arthroscopic bridging reconstruction for irreparable massive rotator cuff tears using autogenous fascia lata were retrospectively analyzed. The age before surgery was 61.3±2.9 years (range 57-67 years). There were 7 patients with right shoulders and 3 with left shoulders. The dominate sides were involved in 7 cases. The trauma history was documented in 2 shoulders. The duration of preoperative symptoms was 14.0±13.5 months (1-48 months). The case with revision surgery was not included. The patients were examined with magnetic resonance imaging (MRI) to evaluate the healing of fascia lata patch bridging in the joint at one week, six months, one year and two years after operation. The motion range of shoulder and the clinical scores, including visual analogue scale (VAS), University of California Los Angeles (UCLA) score, Constant-Murley score and American Shoulder & Elbow Surgeons (ASES) score, were measured before surgery and at follow-up duration. Results: All cases were reconstructed the horizontal couple. No perioperative complication was occurred and all surgery were completed safely and successfully. At the end of two years, the score of ASES was 92.2±3.5 (range 88.3-98.3), UCLA 31.6±2.0 (range 28-34), Constant-Murley 85.2±5.4 (range 78-93) with significant difference (t=11.254, P=0.000; t=12.111, P=0.000; t=8.948, P=0.00) comparing with that before surgery. The VAS pain score was 0.6±0.5 (range 0-1) which was significantly lower than that preoperatively (t=11.326, P=0.000). At 2 years after operation, MRI shows that fascia lata patches healed well in 9 patients. However, one case was with re-tear and patch absorption. The range of motion of shoulder was significantly improved in all patients but with different degrees of weakness (3-4). Conclusion Arthroscopic bridging reconstruction using autogenous fascia lata could effectively improve shoulder function in patients with irreparable massive rotator cuff tears. The autogenous fascia lata patch can heal with the help of rotator cuff tissue through bridging reconstruction.

Objective To investigate the molecular mechanism of colistin resistance mediated by mcr-1and pmrAB genes in clinical Escherichia coli ( E. coli) isolates. Methods A total of 1988 clinical E. coli isolates were collected from the First People′s Hospital of Shangqiu from 2010 to 2017 and screened for colistin-resistant isolates using agar dilution method. The minimum inhibitory concentrations ( MICs) of nine common clinical antibiotics were determined using broth microdilution method. PCR and sequencing analysis were performed to detect the colistin resistance genes of mcr-1 and pmrAB. Conjugation experiments were used to test the transferability of the plasmid carrying mcr-1 gene. S1-PFGE and Southern blot were used to locate the plasmid carrying mcr-1. All colistin-resistant E. coli isolates were typed by multilocus se-quence typing ( MLST) . Results Six colistin-resistant E. coli isolates were obtained by agar dilution meth-od. The results of susceptibility testing showed that all of the six isolates were multidrug resistant. PCR and sequencing analysis revealed that four out of the six strains carried mcr-1 gene, and the other two isolates both had an amino acid substitution (L167P) caused by pmrB gene mutation. The results of conjugation ex-periments, S1-PFGE and Southern hybridization analysis showed that the plasmids of four mcr-1 gene-posi-tive E. coli strains were located on a conjugative plasmid about 60 kb in length. MLST analysis classified the six isolates into six distinct sequence types ( STs) . Conclusion This study suggested that mcr-1 gene and mutations in pmrAB gene were the main mechanisms mediating the resistance of E. coli to colistin. In clinical practice, the occurrence and spread of colistin-resistant E. coli should be further monitored, and the rational use of antibiotics should be promoted to prevent the spread of colistin-resistant strains.