ObjectiveTo investigate the effects of Scutellariae Radix combined with epidermal growth factor receptor-tyrosine kinase inhibitors （EGFR-TKIs） on cell proliferation， apoptosis， cancer stem cell （CSC） marker expression， and metabolism in non-small cell lung cancer （NSCLC） cells. MethodsThe anti-tumor effects of Scutellariae Radix and EGFR-TKIs （gefitinib or osimertinib） in NSCLC cells were evaluated using the cell counting kit-8 （CCK-8） and Annexin V-FITC/propidium iodide （PI） double staining apoptosis assay. The activity of Scutellariae Radix and EGFR-TKIs in three-dimensional （3D） cultures of NSCLC cells was assessed using the CellTiter-Glo® 3D cell viability assay. The mRNA and protein expression levels of CSC markers， sex determining region y box protein 2 （SOX2） and aldehyde dehydrogenase 1 family member A1 （ALDH1A1）， were detected by quantitative real-time polymerase chain reaction （Real-time PCR） and Western blot， respectively. Changes in intracellular reactive oxygen species （ROS） levels were detected by ROS staining， and the redox ratio was detected by femtosecond laser labeling free imaging （FLI）. ResultsUnder both two-dimensional （2D） and 3D culture conditions， compared with the blank group and EGFR-TKI group， the combination group showed significantly reduced cell viability and increased apoptosis rate （P<0.05）. Compared with the EGFR-TKI group， the mRNA and protein levels of CSC markers were significantly downregulated in the combination group （P<0.05）. Additionally， the redox ratio was significantly elevated （P<0.05）， and ROS levels were also increased in the combination group compared with the EGFR-TKI group. ConclusionIn NSCLC cells， Scutellariae Radix enhances the redox ratio and increases ROS levels， thereby inhibiting the expression of CSC markers and strengthening the anti-tumor effects of EGFR-TKIs. This provides a novel molecular mechanism by which Scutellariae Radix may enhance the sensitivity of targeted therapies.

Objective:To explore the effect of moderate-intensity swimming exercise on liver glucose and lipid metabolism disorder in type 2 diabetic rats and possible mechanism. Method:Eight-week-old male SD rats were randomly selected as a blank control group(negative control group,NC group)and fed with normal food,while the rest were fed with high-fat and high-sugar adaptively for one week.Streptozotocin(Streptozotocin,STZ,30mg/kg)was injected intraperitoneally to produce the T2DM rat model.Sixteen rats were randomly selected as type 2 diabetes model group(diabetic model group,DC group)and type 2 diabetes aerobic exercise group(diabetic swimming group,DS group),with eight rats in each group.Aerobic exercise was intervened by non-weight-bearing swimming.After 3 days of adaptive train-ing,the exercise time was 15min/day,and gradually increased to 60min/day,5 days/week,for a total of 8 weeks of training.During the experiment,the body weight,blood sugar,and liver index of rats in each group were detected;ELISA was used to detect the serum insulin(insulin),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β),serum total cholesterol(TC),triglyceride(TG),low-density lipo-protein cholesterol(LDL-C),HDL Lipoprotein cholesterol(HDL-C)and serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels;the expression changes of peroxisome proliferator-activat-ed receptor γ(PPARγ),nuclear factor-κB(NF-κB)and adiponectin(ADPN)in liver tissue were detected. Result:After 8 weeks of aerobic exercise intervention,compared with the DC group,the rats in the DS group had significantly increased body weight,signifiicantly decreased blood sugar(P＜0.01),and significantly increased serum insulin and HDL-C levels(P＜0.01).The serum TNF-α,IL-6,IL-1β and TC,TG,LDL-C,ALT,AST levels were significantly decreased(P＜0.05 or P＜0.01).The liver index and oil red O staining pos-itive area decreased significantly(P＜0.01).The positive area of liver glycogen staining increased significantly(P＜0.01),with neatly arranged liver tissue cells and alleviated fat vacuoles,lipid droplets and inflammatory cell infiltration.The protein and mRNA expression levels of PPARγ,GLUT4 and ADPN in the liver were sig-nificantly increased(P＜0.05 or P＜0.01),and the levels of PPARγ and ADPN immunofluorescence positive cells were significantly increased(P＜0.01).The levels of mRNA and immunofluorescence positive cells of NF-κB were significantly decreased(P＜0.05 or P＜0.01).The mRNA level of ADIPOR2 was also significantly in-creased(P＜0.05),and the protein levels of NF-κB and IL-6 were significantly decreased(P＜0.01). Conclusion:8-week moderate-intensity swimming exercise can improve the disturbance of glucose and lipid me-tabolism by regulating the inflammatory response in the liver of T2DM rats,and the mechanism may be relat-ed to the PPARγ/NF-κB/ADPN pathway.

Objective:To explore the clinical efficacy of free superficial circumflex iliac artery perforator flap (SCIAPF) in reconstruction of soft tissue defects of hand and foot in children.Methods:From August 2021 to August 2023, free SCIAPFs were used to reconstruct soft tissue defects of hands and feet in 8 children at the Department of Hand and Foot Surgery, Zhoukou Huaihai Hospital. There were 6 boys and 2 girls aged between 2 and 7 years old. The sites of soft tissue defect were: 1 of metacarpophalangeal joint in the functional area of left thumb, 2 of dorsal right hand, 1 of dorsal left foot, 2 of medial malleolus of right foot, and 2 of right forefoot, and all were accompanied with tendon and bone exposure. The size of soft tissue defects ranged from 2.5 cm×3.0 cm to 6.0 cm×3.5 cm. After emergency debridement, the children received flap transfers, the flaps sized from 3.0 cm×3.5 cm to 7.0 cm×4.0 cm. Donor sites were closed by running intradermal suture. All children were included in the scheduled postoperative follow-up by visit of outpatient clinic, WeChat interviews and home visits to observe the appearance, colour, texture and sensation of the flaps, the functional recovery of the affected hands and feet, as well as the healing of donor sites.Results:After surgery, all 8 flaps survived beside 1 had a partial necrosis at the edge and healed with a scab after dressing change. Flap donor sites healed in one stage. Scheduled postoperative follow-up lasted from 7 to 22 months, with an average of 10 months. The flaps presented a slightly bloated appearance, soft texture with a restored protective sensation and satisfactory functional recovery of hands and feet. Linear scars were left at the donor sites. According to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, 1 child was of excellent and 2 children of good in hand function of the 3 children with hand injury. The Maryland Foot Score revealed 3 children were of excellent and 2 of good in foot function of the 5 children with foot injury.Conclusion:Free SCIAPF is an ideal option for reconstruction of soft tissue defects of hand and foot in children, with concealed donor site, convenient flap harvesting, fewer complications and satisfactory reconstructive effect.

ObjectiveTo compare and observe the effect of Reduning injection （mainly clearing heat）， Shenfu injection （mainly warming Yang） combined with gefitinib on the proliferation， apoptosis， stemness characteristics and metabolism of lung cancer cells. MethodDifferent non-small cell lung cancer （NSCLC） cell lines were selected and intervened with gefitinib （5， 10， 20 μmol·L^-1）， Reduning injection （0.6%， 0.9%）， Shenfu injection （0.6%， 0.9%）， gefitinib combined with Reduning injection， and gefitinib combined with Shenfu injection. Cell proliferation in each group was detected by cell counting kit-8 （CCK-8） assay， and cell apoptosis was detected by flow cytometry. The mRNA and protein expressions of lung cancer stem cell markers sex determining region Y-box 2 （Sox2） and aldehyde dehydrogenase family 1 member A1 （ALDH1A1） were determind by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The redox ratio of lung cancer cells was observed by femtosecond label-free imaging （FLI） and energy metabolism instrument was used to determine the glycolysis level in cells. ResultCompared with the blank group， Reduning injection reduced the survival rate of lung cancer cells （P<0.05）， increased the apoptosis rate （P<0.05）， down-regulated the mRNA and protein expressions of Sox2 and ALDH1A1 （P<0.05）， and up-regulated the redox ratio of cells （P<0.05）， while Shenfu injection exerted no remarkable effect on the above indexes. In addition， compared with gefitinib alone， Reduning injection combined with gefitinib inhibited the survival rate of lung cancer cells （P<0.05）， promoted the cell apoptosis （P<0.05）， down-regulated the mRNA and protein expressions of Sox2 and ALDH1A1 （P<0.05）， up-regulated the redox ratio of cells （P<0.05）， and lowered the proton efflux rate of glycolysis （P<0.05）， while Shenfu injection combined with gefitinib failed to affect these indexes of lung cancer cells significantly. ConclusionReduning injection may inhibit stemness characteristics of tumor cells by regulating their metabolism to enhance the proliferation-inhibiting and pro-apoptotic effects of gefitinib on lung cancer cells， while Shenfu injection had no significant enhancing effect on gefitinib. This indicates that epidermal growth factor receptor tyrosine kinase inhibitors （EGFR-TKIs） should be used in combination with heat-clearing Chinese medicines.

Objective:To investigate the complement C3a receptor 1 (C3AR1) expression in glioma and its mechanism in progressing malignancy.Methods:(1) The C3AR1 mRNA expression data and clinical information were obtained in 607 glioma patients from The Cancer Genome Atlas (TCGA) database and 656 glioma patients from Chinese Glioma Genome Atlas (CGGA) database; the differences in C3AR1 mRNA expression were analyzed among gliomas with different World Health Organization (WHO) grading. The overall survival and disease-free survival were compared between high and low C3AR1 mRNA expression patients obtained from TCGA database by Gene expression profiling interactive analysis (GEPIA). Gene body (GO) function analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of C3AR1 related differentially expressed genes were performed by DAVID database. Correlation of C3AR1 mRNA expression with immune cell infiltration was analyzed using TIMER online website. (2) The brain tissues from 3 non-tumor patients and 9 glioma patients surgically resected in Department of Neurosurgery, First Affiliated Hospital of Xinxiang Medical University from January 2019 to September 2021 were collected; the C3AR1 protein expression was detected by Western blotting. (3) The in vitro cultured U87 and U251 cells were divided into negative control group and C3AR1 knockdown group ( C3AR1 being knocked down by lentivirus transfection); and CCK-8 assay, plate cloning assay and Transwell assay were used to detect the proliferation rate, number of colony formation and number of membrane penetrating cells. Western blotting was used to detect the nuclear factor-κB (NF-κB) signaling pathway protein expressions. Results:(1) In TCGA database, the C3AR1 mRNA expression in gliomas of WHO grading II, grading III and grading IV increased sequentially, with significant differences ( P<0.05). In CGGA database, the C3AR1 mRNA expression in glioma of WHO grading IV was statistically higher than that in gliomas of WHO grading II and grading III ( P<0.05). GEPIA showed that the overall survival and disease-free survival in the low C3AR1 mRNA expression group were statistically higher than those in the high C3AR1 mRNA expression group ( P<0.05). GO function analysis and KEGG pathway enrichment analysis revealed that C3AR1 related differentially expressed genes were more enriched in such biological processes and signaling pathways as calcium homeostasis, membrane structural valves, proton transmembrane transporter protein activity, chemokine signaling pathway and NF-κB signaling pathway. TIMER showed that C3AR1 mRNA expression in glioblastoma and low-grade glioma was positively correlated with infiltration degrees of B cells, CD4 + T cells, neutrophils, macrophages and dendritic cells, and C3AR1 mRNA expression in glioblastoma was negatively correlated with infiltration degree of CD8 + T cells ( P<0.05). (2) C3AR1 protein expression in glioma tissues was significantly higher than that in non-tumor tissues. (3) Compared with the negative control group, the C3AR1 knockdown group group had significantly lower proliferation rate, smaller numbers of colony formation and membrane penetrating cells, and lower expressions of NF-κB, phosphorylated (p)-NF-κB, p-NF-κB inhibitory protein (IκB)α, p-I-κB kinase (IKK)α and N-cadherin, and significantly higher E-cadherin expression. Conclusion:C3AR1 is highly expressed in glioma and progresses malignancy through NF-κB signaling pathway.

Objective Aimed to develop the evaluation system and weight of hospital orientation training.Methods Literature review,Delphi,questionnaire,AHP to develop the evaluation system and determined the weight with Satty's method.Results The evaluation system includes 3 division's 13 items.Conclusions Course content,teaching method,course difficultness and occupational plan ning play the most important role,and should be paid more attention.

Objective To investigate the effects of autologous blood transfusion and allogeneic blood transfusion on postoperative complications and outcome of patients underwent craniotomy with traumatic brain injury.Methods All transfusional cases underwent emergency craniotomy with trau-matic brain injury from January,2012 to June,201 6,1 61 males and 38 females,ASA physical statusⅠ-Ⅳ,were respectively analyzed and divided into autologous blood group (n = 108)and allogeneic blood group (n =91)based on whether or not using cell salvage.The restrictive transfusion strategy was applied in the two groups and the red blood cells were infused to maintain the hemoglobin concen-tration at 70-100 g/L.The incidence of postoperative complications and adverse transfusion reaction were analyzed and the clinical outcome was judged by Glasgow outcome score (GOS).Results The incidence of postoperative complications (33% vs.56%,P <0.01 )and adverse transfusion reaction (5% vs.14%,P <0.05)of the autologous blood group were lower than that in the allogeneic blood group,and the clinical outcome was better (P <0.01).Logistic regression analysis showed that allo-genetic transfusion (OR =1.953,95%CI 1.381-2.529)was an independent risk factor of postopera-tive complications.Conclusion The use of autologous blood transfusion in patients with traumatic brain injury can reduce the incidence of postoperative complications and the risk of blood transfusion and improve clinical outcome.

Objective To analyze the changes of serum levels ofαII-spectrin breakdown products (SBDPs)in traumatic brain inj ury (TBI)patients,and further to investigate the clinical diagnosis value of SBDPs for patients with TBI,especially with mTBI.Methods The serum levels of SBDPs were examined in 43 severe TBI (sTBI)patients,43 mild TBI (mTBI)patients and 43 healthy controls using enzyme linked immunosorbent assay (ELISA).The diagnostic usefulness of SBDPs for TBI patients were assessed by Receiver Operating Characteristic (ROC)curves analysis.Results There was no significant difference of SBDP145 among the three groups (F=1.340,P>0.05).Serum levels of SBDP120 in controls,mTBI and con-trols were 7.06±2.23,11.67±9.14 and 12.64±11.44 ng/ml,respectively.Compared with controls,serum levels of SB-DP120 were significantly higher in patients with sTBI (F=9.873,P=0.001)and mTBI (F=9.873,P=0.008),while there was no significant difference of SBDP120 between sTBI patients and mTBI patients (F=9.873,P=0.515>0.05). The area under ROC curve (AUC)of SBDP120 for TBI patients was 0.781 (95% CI:0.690~0.872,P<0.001).For mTBI patients,the area under ROC curve was 0.736 (95% CI:0.624~0.848,P<0.001).And for discriminating TBI patients with CT negative or positive,the area under ROC curve was 0.709 (95% CI:0.582~0.837,P=0.007<0.01).Conclusion The serum levels of SBDP120 were significantly increased in TBI patients,especially mTBI patients.And the serum levels of SBDP120 can be used as potential non-invasive biomarker for mTBI patients.

Objective To detect the expression of micro RNA (miR)-433 and histone deacetylase 6 (HDAC6) in glioma tissues and investigate the effect of miR-433 on cell proliferation and invasion of human glioma cell line U251. Methods (1) Forty-two glioma samples, collected from patients accepted surgical resection and conformed by pathology in our hospital from January 2010 to December 2014, and 13 healthy brain tissues, collected from patients accepted surgery for craniocerebral trauma at the same time period, were used in our study; reverse transcription (RT)-PCR was used to detect the mRNA expression levels of miRNA-433 and HDAC6 in the glioma samples and brain tissues. (2) Human glioma cell line was routinely cultured and divided into blank control group, nonsense sequence control group and miRNA-433 mimics group;cells in the later two groups were transfected with nonsense sequences or miRNA-433 mimics, and cells in the blank control group did not give any treatment;the mRNA expression levels of miRNA-433, P21 and HDA C6 in these 3 groups were detected by RT-PCR;the cellular viability was measured by CCK-8 assay;flow cytometry was used to monitor the changes of cell cycle and apoptosis; cell invasion was evaluated by Transwell assay; HDAC6 protein expression was detected by Western blotting. (3) Wide-type (WT)HDAC63'-UTR and mutant type (MUT)HDAC63'-UTR luciferase report vectors were established; miR-433 mimics+WT HDAC63′-UTR and nonsense sequences+WT HDAC63'-UTR were transfected into the U251 cells, and dual-luciferase experiment was used to detect the fluorescence intensity of the cells; miR-433 mimics+MUT HDAC63'-UTR and nonsense sequences+MUT HDAC63'-UTR were transfected into the U251 cells, and dual-luciferase experiment was used to detect the fluorescence intensity of the cells. (4) U251 cells were divided into nonsense sequence control group, HDAC6 expression plasmids group and HDAC6 siRNA group, and nonsense sequences, HDAC6 expression plasmids or HDAC6 siRNA were transfected respectively; RT-PCR was used to detect the P21 and HDAC6 mRNA expressions and miRNA-433 expression; U251 cells were divided into miR-433 mimics group and miR-433 mimics+HDAC6 expression plasmids group, and miR-433 mimics or miR-433 mimics+HDAC6 expression plasmids were transfected, respectively, and one-5 d after that, CCK-8 was used to detect the cellular viability. Results (1) The miRNA-433 expressions gradually increased and HDA C6 mRNA expressions gradually decreased in the high-grade gliomas, low-grade gliomas and normal brain tissues, and significant differences were noted among each two groups (P<0.05);the miRNA-433 expression was negatively correlated with HDA C6 mRNA expression in the glioma tissues (r=0.829, P=0.000). (2) As compared with blank control group and nonsense sequence control group, miRNA-433 mimics group had significantly higher miRNA-433 and P21 mRNA expressions, cell percentage at G0/G1 stage, and apoptotic rate (P<0.05), and had statistically lower HDAC6 mRNA expression, cellular viability on 2-5 d of culture, number of transmembrane cells and HDAC6 protein expression (P<0.05). (3) The luciferase activity in cells from miR-433 mimics+WT HDAC63'-UTR group was significantly lower as compared with that in the nonsense sequences+WT HDAC63'-UTR group (P<0.05);the luciferase activity in cells from miR-433 mimics+MUT HDAC63'-UTR group and nonsense sequences+MUT HDAC63'-UTR group showed no significant differences (P>0.05). (4) The HDA C6 mRNA expressions were gradually increased, and P21 mRNA expressions were gradually decreased in the HDAC6 siRNA group, nonsense sequence control group, and HDAC6 expression plasmids group, with significant differences (P<0.05);on 2-5 d of culture, the cellular viability in the miR-433 mimics+HDAC6 expression plasmids group was significantly higher than that in the miR-433 mimics group (P<0.05). Conclusions The miRNA-433 expression level is low in human glioma tissues;miRNA-433 over-expression may inhibit the cell activity and promote cell apoptosis of glioma cell line U251 in vitro via inhibiting the HDAC6 expression.

Objective To explore the effects of ultrasound-exposed microbubbles (UM) on the expression of CXC chemokine receptor 4 (CXCR4) in bone marrow mesenchymal stem cells (BMSCs) and the mechanisms involved.Methods Mesenchymal stem cells were isolated from bone marrow taken from male Sprague-Dawley rats.They were divided into a control group,an ultrasound (US) group,an ultrasound-exposed microbubbles (UM) group,a UM plus catalase (UMC) group,a UM plus AMD3100 (UMA) group,and a UM plus anti-CXCR4 antibody (UMCX) group.The control group was not given any treatment.The US group was treated with 1 MHz ultrasound at 1 Watt per square centimetre for 30 seconds.The UM group was treated with ultrasound plus microbubbles.The UMC group was treated with catalase,microbubbles and ultrasound.The UMA group was treated with AMD3100,microbubbles and ultrasound.The UMCX group was treated with anti-CXCR4 antibody,microbubbles and ultrasound.Quantitative polymerase chain reaction (qPCR) and Western blotting were performed to determine the levels of CXCR4 mRNA transcription and the expression of BMSCs in the control,US,UM and UMC groups.Immediately,5 minutes and 15 minutes after the intervention,fluorescence intensities were observed in the cells labeled with Fluo-4/AM of the control group,US group and UM group under a fluorescence microscope.Migration assays were conducted to determine the chemotactic ability of the BMSCs with respect to stromal-derived factor-1α (SDF-1α) in all six groups.Results No significant differences were found in the levels of CXCR4 mRNA transcription and protein expression between the US and control groups(P＞0.05),but the levels in those groups and the UMC group were lower than those observed in the UM group.Fluorescence intensity in the cells of the US group was not significantly different from that in the control group (P＞0.05),but those levels were both significantly lower than that in the UM group (P＜0.05).There was no significant difference in the number of cells migrating to the SDF-1α between the US (22.4±2.2) and control group (20.5±2.3).However,the number of cells migrating to SDF-1α in the UM group (53.1±3.8) was significantly larger than that in the US group,the control group,the UMC group (35.2+3.1),the UMA group (32.5±2.8) and the UMCX group (30.7+2.9) (P＜ 0.05).Conclusion UM can increase mRNA transcription and the expression of CXCR4 protein in BMSCs,and promote BMSCs migration to SDF-lα.This may in part be mediated by an increase in calcium influx.