Anlotinib has strong antiangiogenic effects and leads to vessel normalization.However,the"window period"characteristic in regulating vessel normalization by anlotinib cannot fully explain the long-term survival benefits achieved through combining it with other drugs.In this study,through RNA sequencing(RNA-seq)and label-free quantitative proteomics analysis,we discovered that anlotinib regulated the expression of components of the extracellular matrix(ECM),leading to a significant reduction in ECM stiffness.Our bioinformatic analysis revealed a potential positive relationship between the ECM pathway and gefitinib resistance,poor treatment outcomes for programmed death 1(PD-1)targeting,and unfavourable prognosis following chemotherapy in lung cancer patients.We administered anlotinib in combination with these antitumour drugs and visualized their distribution using fluorescent labelling in various tumour types.Notably,our results demonstrated that anlotinib prolonged the retention time and distribution of antitumour drugs at the tumour site.Moreover,the combination therapy induced notable loosening of the tumour tissue structure.This reduction was associated with decreased interstitial fluid pressure and tumour solid pressure.Additionally,we observed that anlotinib effectively suppressed the Ras homologue family member A(RhoA)/Rho-associated protein kinase(ROCK)signalling pathway.These findings suggest that,in addition to its antiangiogenic and vessel normalization effects,anlotinib can increase the distribution and retention of antitumour drugs in tumours by modulating ECM expression and physical properties through the RhoA/ROCK signalling pathway.These valuable insights contribute to the development of combination therapies aimed at improving tumour targeting in cancer treatment.

Objective To construct models of viral transfection and hypoxia/reoxygenation induced cellular injury in adult mouse cardiomyocytes(AMCMs)isolated using a non-Langendorff method.Methods AMCMs were isolated,extracted,sedimented,and plated using a non-Langendorff method.The morphology and survival rate of the isolated cells were evaluated 2,24,48 and 72 h after plating,and their integrity was observed by immunofluorescence staining for α-actinin.The isolated AMCMs were infected with adenoviruses carrying an RFP-expressing vector and fluorescence images were obtained at 36 and 48 h post-infection and used to calculate transfection efficiency.The cells were cultured under hypoxic conditions for 45 min,reoxygenated for 24 h,and then stained with propidium iodide(PI)to verify establishment of the hypoxia/reoxygenation injury model.Results The survival rates of AMCMs at 2,24 and 48 h after plating were comparable,but survival was significantly reduced at 72 h.The integrity of the AMCMs was good and＞80%of the cells were transfected with adenovirus at 48 h.After hypoxia/reoxygenation treatment,42%of cells were stained by PI,suggesting successful establishment of the AMCM injury model.Conclusions In this study,we developed a non-Langendorff method for the fast and easy isolation of AMCMs with high cell viability.The isolated cells can be efficiently infected with adenovirus and respond to hypoxia/reoxygenation injury.These findings provide a systematic method for isolating AMCMs and for applying gene modification and hypoxia/reoxygenation injury in these cells.

Objective:To compare the efficacy and safety of azacitidine combined with venetoclax or CAG regimen in the treatment of newly treated elderly patients with acute myeloid leukemia (AML).Methods:A retrospective cohort study was conducted. The clinical data of 34 newly treated elderly patients with AML treated in the Fifth People's Hospital of Datong from May 2018 to August 2023 were retrospectively analyzed. According to the treatment regimen, all patients were divided into venetoclax group (azacitidine + venetoclax, 17 cases) and CAG group (azacitidine + CAG regimen, 17 cases). The clinicopathological characteristics, efficacy, adverse reactions and survival of the both groups were compared.Results:There were no statistically significant differences in the clinical data of both groups (all P > 0.05). The complete remission (CR) rate and the objective response rate (ORR) in venetoclax group were higher than those in CAG group [CR: 70.6%(12/17) vs. 47.1% (8/17); ORR: 82.4% (14/17) vs. 64.7% (11/17)],while the differences in CR and ORR were not statistically significant (χ 2 = 2.00, P = 0.163; χ 2 = 2.00, P = 0.244). The follow-up time[ M ( Q1, Q3)] was 25.4 months (7.2 months, 60.3 months). At the end of follow-up, 19 of 34 patients survived (13 cases in venetoclax group and 6 cases in CAG group); 15 died (4 cases in venetoclax group and 11 cases in CAG group). The median overall survival (OS) time was 14.22 months (95% CI: 8.2-60.3 months) and 10.56 months (95% CI: 7.2-50.2 months), respectively in venetoclax group and CAG group;the median progression-free survival (PFS) time was 9.97 months (95% CI: 5.4-40.5 months) and 6.82 months (95% CI: 5.0-36.2 months), respectively, and there were no statistically significant differences in OS and PFS between the two groups (all P > 0.05). Grade 3-4 hematological adverse reactions occurred in 16 and 14 patients in venetoclax group and CAG group, respectively. There were no significant differences in granulocyte deficiency time, platelet deficiency time, infection and bleeding incidence between the two groups (all P > 0.05). Conclusions:Azacitidine combined with venetoclax or CAG regimen have better clinical efficacy and safety for newly treated elderly patients with AML.

Objective:To investigate the effect and adverse reactions of radiotherapy combined with targeted drugs for the treatment of multiple brain metastases (MBM) in patients with non-small cell lung cancer (NSCLC) and the changes in serum tumor marker levels.Methods:A retrospective cohort study was conducted. Eighty-six patients with NSCLC-MBM who were admitted to the Fifth People's Hospital of Datong from June 2019 to June 2022 were selected, and the patients were divided into the study group and the control group according to different treatment methods, with 43 cases in each group. The study group was given radiotherapy to the tumor primary focus combined with erlotinib or gefitinib targeted therapy, and the control group was given radiotherapy to the tumor primary focus based on conventional chemotherapy with pemetrexed combined with platinum-based drugs. The efficacy, overall survival rate and incidence of adverse reactions were compared between the two groups at 1 month after treatment; the levels of serum tumor markers S100-β, carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCCA) were compared between the two groups before and after treatment.Results:The study group was aged (55±5) years old, ranging from 39 to 75 years old, including 15 (34.88%) males and 28 (65.12%) females; the control group was aged (54±5) years old, ranging from 38 to 72 years old, including 17 (39.53%) males and 26 (60.47%) females. The general data such as age and gender of patients were compared between the two groups, and the differences were not statistically significant (all P>0.05). The objective remission rate [53.49% (23/43) vs. 32.56% (14/43)] and disease control rate [93.02% (40/43) vs. 69.77% (30/43)] at 1 month after treatment, and 1-year overall survival rate [58.14% (25/43) vs. 46.51% (20/43)] of the study group were higher than those of the control group, and the differences were statistically significant ( χ2 values were 10.91, 5.76 and 11.02, respectively; P values were 0.001, 0.016 and 0.001, respectively). Before treatment, the differences in serum S100-β, CEA and SCCA levels between the two groups of patients were not statistically significant (all P > 0.05). At 1 month after treatment, the serum S100-β, CEA and SCCA levels of the study group were lower than those of the control group, and the differences were statistically significant (all P < 0.05). The proportion of patients with adverse reactions in the study group was lower than that in the control group [13.95% (6/43) vs. 39.53% (17/43)], and the difference was statistically significant ( χ2 = 8.35, P < 0.05). Conclusions:Radiotherapy combined with targeted drugs therapy may prolong the survival of NSCLC-MBM patients, reduce the occurrence of adverse reactions and decrease the levels of serum tumor markers, which is worthy of clinical promotion.

Objective:To compare the efficacy and safety of azacitidine alone or combined with half-course CAG (arorubicin + cytarabine + granulocyte colony stimulating factor) regimen and azacitidine combined with full-course CAG regimen in treatment of patients with acute myeloid leukemia (AML) or myelodysplastic syndromes (MDS).Methods:The clinical data of 51 patients with AML and MDS admitted to Datong Fifth People's Hospital from September 2019 to March 2022 were retrospectively analyzed. Among them, 17 patients received azacitidine alone 7-day regimen, 17 patients received azacitidine combined with half-course CAG regimen and 17 patients received azacitidine combined with full-course CAG regimen. The remission rate, adverse reaction rate and supportive treatment were compared among the three groups.Results:The objective remission rate (ORR) was 58.8% (10/17), 64.7% (11/17) and 70.6% (12/17) in azacitidine alone group, azacitidine combined with half course CAG group, and azacitidine combined with full course CAG group, respectively, and the difference was not statistically significant among the above groups ( P = 0.773). The main adverse reaction after treatment with azacitidine was bone marrow suppression,and 32 patients had grade 3-4 hematological side effects. The average time of agranulocytopenia was (15±5) d, 23 patients had infection and 11 cases had hemorrhage. There were no significant differences of the three groups in the hemorrhage incidence, the infection, incidence, agranulocytosis time, the amount of red blood cell infusion and the amount of platelet infusion (all P > 0.05). Except 1 patient died of acute left ventricular dysfunction after chemotherapy in the first cycle and 1 patient died of cerebral hemorrhage after chemotherapy in the third cycle, all the patients successfully completed the chemotherapy after active symptomatic support treatment and safely passed the bone marrow suppression period. Conclusions:Azacitidine alone, azacitidine combined with half-course CAG, azacitidine combined with full-course CAG regimens in the treatment of AML/MDS all show good curative effects, and their adverse reactions are similar to supportive treatment. Azacitidine combined with full-course CAG regimen has a relatively high effective rate.

Objective:To explore the performance of the commonly used whole blood C-reactive protein (CRP) detection systems and give related recommendation on the performance requirements of detection systems.Methods:A total of 7 540 venous blood samples from 26 maternal, child and children′s hospitals were collected to conduct this multi-center study on the analytical performance of 5 commonly used whole blood CRP detection systems from March to April in 2019. The blank check, carryover, repeatability, intermediate precision, linearity, sample stability, influence of hematocrit/triglyceride/bilirubin, comparison with SIEMENS specific protein analyzer and trueness were evaluated. The 5 systems included BC-5390CRP autohematology analyzer, AstepPLUS specific protein analyzer, Ottoman-1000 Automated Specific Protein POCT Workstation, i-CHROMA Immunofluorometer equipment Reader and Orion QuikRead go detecting instrument. The 5 systems were labeled as a, b, c, d and e randomly.Results:Within the 5 systems, all values of blank check were less than 1.00 mg/L, the carryovers were lower than 1.00%. The repeatability of different ranges of CRP concentrations including 3.00-10.00, 10.00-30.00 and>30.00 mg/L were less than 10.00%, 6.00% and 5.00%, respectively, and the intermediate precision was less than 10.00%. The linearity correlation coefficients of the 5 systems were all above 0.975, while the slope was within 0.950-1.050. Whole blood samples were stable within 72 hours both at room temperature (18-25 ℃) and refrigerated temperature (2-8 ℃). The CRP results were rarely influenced by high triglyceride or bilirubin, except for the immmunoturbidimetric test based on microparticles coated with anti-human CRP F(ab) 2 fragments. When triglyceride was less than 15.46 mmol/L, the deviation of CRP was less than 10.00%. When bilirubin was less than 345.47 μmol/L, the deviation of CRP was less than 10.00%. CRP was more susceptible to Hct on the systems without Hct correction. The deviation of CRP between different Hct dilution concentration and 40% dilution concentration can reach as high as 67.48%. The correlation coefficients ( r) of 5 systems were all more than 0.975 in the range of 0-300.00 mg/L compared with Siemens specific protein analyzer. All systems passed the trueness verification using the samples with specified values of 12.89 and 30.60 mg/L. Conclusion:The performance of 5 systems can basically meet the clinical needs, but it is suggested that the whole blood CRP detection system without automatic Hct correction should be modified manually.

Objective To evaluate the clinical outcomes and correction accuracy of deformities around the knee joint using external fixator-assisted internal fixation (FAIF).Methods From January 2014 to March 2018,data of 15 patients (17 bone segments) with deformities around the knee joint who had been corrected with external fixator-assisted internal fixation were retrospectively analyzed.There were 6 males and 9 females,with an average age of 39 years (range,21-60 years).There were 9 patients with genu varus and 6 with genu valgus.All the 15 patients had shortening ranging from 5 to 22 mm,with an average of 12 mm.Unilateral external fixator was used to stabilize the osteotomy site intraoperatively until the satisfied mechanical axis was acquired.Then the plate or Intramedullary nail was inserted and fixed,the external fixator was removed after internal fixation.Results Femur osteotomy was done in 7 patients and tibia osteotomy was done in 6 patients,while 2 patients had osteotomy in both tibia and femur.Intramedullary nailing was used in 2 bone segments,and plate was used in 15 bone segments.The correction of angle ranged from 7° to 22°,with an average of 12.5°.The correction of the 9 cases of genua varus ranged from 10° to 22°,with an average of 17.6°.The correction of the 6 cases of genu valgum ranged from 7° to 13°,with an average of 10.4°.The mean time to achieve union of the osteotomy sites was 3.5 months (range,2.5-5.0 months).All the 15 patients were followed up for 8-48 months (average,23 months).The mechanical axis deviation (MAD) was 5.93 mm (range,0-15 mm) after operation which was 34.8 mm (range,8-55 mm) before operation.The mean post-operative mechanical lateral distal femoral angle (mLDFA) was 87.5° (range,84°-90°) which was 76.1° (range,66°-82°) before operation.The mean medial proximal tibia angle (MPTA) was 87.8° (range,86°-89°) which was 76.8° (range,68°-80°) before operation.There were no deep infection and neurovascular injury.Conclusion The FAIF not only has the advantage of minimal invasiveness and easy adjustment,but also can avoid uncomfortableness and complications of long-term of wearing the external fixator.It is an accurate and safe method to correct the deformities around the knee joint.

Objective To explore the predictive value of Tei index and plasma brain natriuretic peptide(BNP) on major adverse cardiovascular events(MACE) during long term follow up in the patients with anteriormyocardial infarction(MI).Methods A total of 238 patients with anterior MI were selected and divided into four groups according to the median of Tei index and BNP:G1 group(Tei≤0.66,BNP≤532.60 ng/mL,n=70),G2 group(Tei≤0.66,BNP＞532.60 ng/mL,n=51),G3 group(Tei＞0.66,BNP≤532.60 ng/mL,n=50) and G4 group(Tei＞0.66,BNP＞532.60 ng/mL,n=67).The follow up end was the MACE occurrence.Results There was a significant positive correlation between the Tei index and BNP(r=0.582,P=0.000).Tei entered the BNP regression equation(β=0.658,P＜0.05).The MACE-free survival rate had no statistical difference among 4 groups(x2 =9.975,P=0.019).The Cox regression analysis showed that Tei index and BNP were the independent predictive factors for MACE.Tei combined with BNP had stronger predictive value to MACE(AUC=0.781,95CI:0.721-0.841,P=0.000),its sensitivity was 72.52％ and specificity was 76.89％.Conclusion Tei combined with BNP has stronger predictive value for anterior MI during long term follow up.

Objective To retrospectively summarize the diagnosis and treatment experience of acute renal failure caused by urinary calculi obstruction in infants under the age of 6 months in order to improve the level of treat-ment. Methods Between January 2010 and December 2014,15 infants under the age of 6 months with bilateral urinary tract calculi obstruction leading to acute renal failure were treated in West China Hospital,Sichuan University. All cases were Tibetan(10 males,5 females). The ages ranged from 1 month 15 days to 5 months 24 days. The weights ranged from 3. 5 to 7. 0 kg. The clinical characteristics,treatment and clinical outcome were analyzed. Results A total of 9 infants(5 males,4 females)were assessed to be able to tolerate general anesthesia surgery and received positive conservative treatment before operation,and obstruction relief within 6 hours of admission. Six infants (5 males, 1 female)with serious internal milieu disorder,pneumonia,and shock,were assessed as critical cases,who could not tolerate general anesthesia surgery temporarily. Under the emergency blood purification support,they received anti -infection,spasmolysis,rehydration,to correct the internal milieu disorder and obstruction relief within 12 hours of admis-sion. Serum potassium,creatinine returned to normal 12 - 48 hours after obstruction relief. Stone specimens were obtained from 5 cases,and the stone compositions were calcium oxalate stone in 3 cases,calcium oxalate and carbonate apatite mixed stone in 2 cases. A total of 13 cases need further urolithiasis treatment. Conclusions The infants under the age of 6 months with bilateral urinary tract calculi obstruction are more severe with more complications. The emergency treatment principle is to save lives,then relieve the obstruction;surgery should be as brief as possible,since removing the stones is not the primary purpose. If the infants are in critical,life - threatening circumstances,and cannot tolerate general anesthesia surgery,the blood purification will be helpful to gain time and create conditions for treatment.

Objective: To study the relationship between insulin resistance (IR) and coronary collateral circulation in patients with impaired glucose tolerance (IGT). Methods: A total of 227 patients with coronary angiography (CAG) were studied. There were 131 patients with male gender and the average patient’s age was (53.2 ± 11.0) years. IR (HOMA2-IR) index was measured by HOMA2 method, the severity level of coronary stenosis was assessed by Gensini scoring system, collateral circulation condition was determined by Rentrop classiifcation. 187 IGT patients were divided into 4 groups: Rentrop 0 group,n=55, Rentrop 1 group,n=42, Rentrop 2 group,n=39 and Rentrop 3 group,n=51; in addition, Control group,n=40 patients with normal glucose tolerance and coronary stenosis<50%. Results: Compared with Control group, all patients in 4 Rentrop groups had increased 2h-PBG, HbA1c, HOMA2-IR and Gensini score, while decreased fasting insulin (FINS), allP<0.05. Compared with Rentrop 3 group and Rentrop 2 group, the patients in Rentrop 1 group and Rentrop 0 group had elevated 2h-PBG, HbA1c, HOMA2-IR and Gensini score, while Rentrop 0 group had reduced FINS, allP<0.05. Multivariable regression analysis showed that HOMA2-IR index (R=0.518,P<0.05), HbA1c (R=1.916, P<0.05), 2h-PBG (R=2.130,P<0.05) and FINS (R=1.547,P<0.05) might be related to the severity of coronary stenosis. Binary regression analysis indicated that poor collateral circulation (the patients in Rentrop 0 group and Rentrop 1 group) was related to HOMA2-IR index (OR=1.679, 95% CI 1.101-2.558,P=0.016). Conclusion: HOMA2-IR index could be signiifcantly higher in patients with IGT combining chronic coronary occlusion. IR was the independent risk factor for the severity of coronary stenosis and coronary collateral formation.