ObjectiveTo understand the characteristics of mutations in BCR-ABL1 kinase domain mutation,these chronic myeloid leukemia (CML) and Ph positive acute lymphoblastic leukemia (ALL)patients who got imatinib treatment had poor effect.MethodsTotally 177 CML patients and 33 Ph( + )ALL patients were selected at Beijing Dao-Pei Hospital from Sep.2007 to Dec.2010.All of them were Chinese patients.Totally 243 bone marrow or peripheral blood specimens were collected from the patients,who had early effect,then resistance emergenced,or for more than 3 months of poor efficacy.Extracted total RNA from the specimens' nuclear cells,reversed transcription to cDNA.Amplified the whole span of BCRABL1 fusion kinase gene by nest PCR (from 242 to 493 amino acid coding sequence),used the type AB3130XL gene sequencing instrument determinate the gene sequence of ABL1 kinase region and then used the Variant Reporter V1.0 software to analyze the results of gene mutations.ResultsThirty-two kinds of different mutations were detected of ABL1 gene mutations,accounting for 34.2％ (83/243 cases).Among them,the T315I was 12％ (10/83),mutation rate was the highest,followed by Y253H was 11％ (9/83),G250E was 7％ (6/83),E255K was 7％ (6/83),M351T was 6％ (5/83),E459K was 5％ (4/83) ;Q252H,D276G,F317L,E355G,F359V,H396R were all 4％ (3/83).Three cases of insertion mutations were found,including 2 cases of 357-358insk,1 case of V304RfsX17.Seven patients had found existence two or more point mutations.The multiple drug resistance mutations might exist in the same leukemia clone.The same individual was not only contain common resistance mutations,but also rare point mutations,insertion mutations.The mutations might be lead to loss of kinase activity.ConclusionsUnder the imatinib drugs pressure,the ABL1 gene mutation in leukemia cells appears randomly,and results in different resistant clones.Different resistant clones can coexist in the same patients in vivo; resistant clones not only contain point mutations,but also contain inserted deletion mutations.

AIM:To observe the expression of tyrosine hydroxylase (TH), ionized calcium binding adapter molecule 1 (Iba1) and pro-opiomelanocortin (POMC) in rat arcuate nucleus (Arc) of hypothalamus induced by substantia nigra (SN) lesion with 6-hydroxydopamine (6-OHDA).METHODS:According to the random number table, 20 SD rats were randomly divided into 6-OHDA group and control group.6-OHDA was injected into the bilateral SN of the rats in 6-OHDA group, and the same volume of saline was injected into the same position of the rats in control group.After 6 weeks, the rats were sacrificed and the brains were obtained.Immunofluorescence staining and Western blot were used to detect TH expression in the SN, and TH, Iba1 and POMC expression in the Arc.RESULTS:Compared with control group, no significant change of body weight in 6-OHDA group was observed, but the weight of retroperitoneal fat decreased from (7.550±0.670) g to (3.895±0.465) g (P<0.01).The number of TH immunoreactive neurons in SN decreased from 56±5 to 14±2 (P<0.05), and the TH protein level decreased from 0.75±0.11 to 0.41±0.09 (P<0.01).In the Arc, the number of TH immunoreactive neurons decreased from 21±2 to 10±3 (P<0.05), the number of Iba1 immunoreactive neurons increased from 12±2 to 30±5 (P<0.05), and the number of POMC immunoreactive neurons increased from 16±4 to 31±2 (P<0.05).In the hypothalamus, the TH protein level decreased from 0.35±0.05 to 0.21±0.02 (P<0.01), the Iba1 protein level increased from 0.23±0.06 to 0.51±0.04 (P<0.01), and the POMC protein level increased from 0.37±0.05 to 0.65±0.03 (P<0.01).CONCLUSION:The changes of TH, Iba1 and POMC expression in Arc of 6-OHDA-treated rats may involved in the fat loss in Parkinson's disease.

Objective To analyze the correlative factors of anemia in patients with non-small-cell lung cancer (NSCLC), and to explore the impact of anemia on prognosis.Methods The clinical data of 473 patients with NSCLC treated at the first time from January 2008 to November 2012 in the Fourth Hospital of Hebei Medical University were analyzed retrospectively.Results There were 273 patients (57.72 ％) with anemia.Anemia occurred in different age (x2 =3.459, P =3.459), different albumin level (x2 =70.648, P =70.648), different PS score (x2 =10.222, P =10.222), whether recent surgery (x2 =4.956, P =4.956), whether recent chemotherapy (x2 =3.627, P =0.037), and other factors.By multiple factors analysis, hypoalbuminemia was an independent risk factor for anemia (P ＜ 0.05).The median OS of the anemia patients was shorter than that of the patients without anemia (15 months vs 17 months, P ＜ 0.05).Conclusions Hypoalbuminemia is the independent risk factor for emergence of anemia.Anemia is the prognosis indicator of shorten survival period, which is an independent factor of prognosis in the NSCLC.

ObjectiveTo assess the effects of constraint-induced movement therapy(CIMT) on sub-acute stroke patients with upper extremity motor dysfunction.Methods63 patients with hemiplegia after stroke were divided into two groups: control group(31 cases) and CIMT group(32 casese). The two groups received the regular rehabilitation training for 14 d. Then the control group went on the the regular rehabilitation training, while the CIMT group received CIMT for 14 d. All patients were assessed by the Action Research Arm Test(ARAT) and Fugl-Meyer(FMA) on the first day, the 15th day and the 30th day after the treatment.ResultsARAT score and FMA scores were higher in CIMT group than in the control group 30 d after the treatment and in CIMT group 15 days after the treatment(P<0.001).ConclusionCIMT is more effective to improve the upper limb motor function of the sub-acute stroke patients than the regular rehabilitation training.

Objective The present study aimed to explore the effect of miR-29b on ox-LDL-stimulated vascular smooth muscle cells and the underlying mechanism.Methods VSMCs were divided into four groups:control+NC group,control+miR-29b group,ox-LDL+NC group and ox-LDL+miR-29b group.The cells were treated with 80 mg/L ox-LDL for 24 h to estab-lish the model.A cell counting Kit-8(CCK-8)assay was performed to evaluate VSMC proliferation.The expression of miR-29b,MCP-1,MCP-3,SM α-actin,SM MHC and KLF4 was detected via quantitative reverse-transcriptase polymerase chain reaction(qRT-PCR).The expression of MCP-1 and MCP-3 in the supernatant was detected via ELISA.Protein expression was exam-ined via Western blotting.A luciferase reporter assay was used to identify the relationship between KLF4 and miR-29b.Results We found that miR-29b was downregulated significantly in ox-LDL-stimulated VSMCs.The miR-29b mimics inhibited the pro-liferation and expression of KLF4,MCP-1 and MCP-3 and increased the levels of SM α-actin and SM MHC in the VSMCs trea-ted with ox-LDL.miR-29b could bind to the 3'-untranslated region(3'UTR)of KLF4 mRNA and subsequently inhibit the activi-ty of luciferase.The overexpression of KLF4 abrogated the effects of the miR-29b mimics.Conclusion In summary,miR-29b is able to regulate the proliferation and contractile markers of VSMCs via directly regulating KLF4.

Extracellular vesicles are a kind of membrane structures deriving from cells, which exist widely in urine, blood, saliva, milk, cerebrospinal fluid, amniotic fluid and other body fluid.Extracellular vesicles contain nucleic acid, enzymes, proteins and other bioactive molecules, which can convey the biological information to target cells through a variety of modes and participate in the pathophysiological processes of target cells.In recent years, it is considered that extracellular vesicles are associated with the pathogenesis of inflammatory disease of pulmonary such as asthma, chronic obstructive pulmonary disease and acute lung injury/acute respiratory distress syndrome, and have potential therapeutic value through a large body of research.

Background and objective hTe activation of coagulation and ifbrinolysis is frequently encountered among cancer patients. Such tumors are associated with high risk of invasion, metastases, and negative ifnal outcomes. Non-small cell lung cancer (NSCLC) accounts for approximately 80%to 85%of all lung malignancies. hTis study aims to investigate the prognostic value of blood coagulation tests for NSCLC and provide a reference to patients on the prevention and treatment of thrombophilia. Methods Data were collected from 604 cases of hospitalized patients with histologically conifrmed NSCLC from January 2009 to December 2012 at the Fourth Hospital of Hebei Medical University. Data included the related indexes of coagulation function in patients before treatment [(i.e., prothrombin time (PT), prothrombin time activity (PTA), interna-tional normalized ratio (INR), activated partial thromboplastin time (APTT), ifbrinogen (Fib), D-dimer, and platelet count)], as well as sex, age, pathological type, TNM stage, and lymph node status. Fitfy control subjects without cancer were included in the analysis. Statistical analysis was conducted by using SPSS 13.0 sotfware. Results hTe plasma level of all coagulation tests including D-dimer, Fib, PT, APTT, INR, and platelet counts revealed statistically signiifcant differences between the patient and control group (P<0.001 for all variables;P=0.001,5 and P=0.004,5 for Fib and platelet counts, respectively). hTe squamous subtype exhibited high plasma Fib levels (P<0.001) compared with adenocarcinoma cell lung cancer patients. Fib and PLT levels increased (P<0.001 and P=0.014, respectively), and aPTT decreased (P<0.001) in patients at stages III and IV compared with those in patients at stages I and II. aPTT decreased signiifcantly (P<0.001), and Fib and D-dimer levels increased (P<0.001 and P=0.048, respectively) in N1-3 patients with NSCLC compared with those of N0 patients. Prolonged PT and INR, high plasma Fib levels, and low PTA levels had statistically signiifcant adverse effects on survival (P=0.032, P=0.001, P<0.001, and P=0.005, respectively). Multivariate analyses revealed that only increased INR was associated with a decreased survival rate in the related indexes of coagulation function (P=0.017). Conclusion Patients who have lung adenocarcinoma and/or lymph node metastasis at advanced stages of NSCLC are prone to thrombophilia. Prolonged PT and INR was associated with poor prognosis, and elevated INR was independently associated with a decreased survival rate. PT and INR are promising prognos-tic markers of NSCLC.

Objective:To investigate the relationship between serum matrix metalloproteinase-9 (MMP-9) level and the location and severity of bleeding in patients with cerebral microbleeds(CMBs).Methods:A total of 60 CMBs patients admitted to the Department of Neurology of the First Affiliated Hospital of the Xinxiang Medical University from January 2019 to August 2020 were selected as subjects as the CMBs group, and 60 healthy controls without nervous system diseases in outpatient physical examination during the same period were selected as the control group. The clinical data and biochemical indicators of the two groups were collected. Serum MMP-9 levels were measured by enzyme linked immunosorbent assay (ELISA). According to susceptibility weighted imaging (SWI), CMBs patients were divided into grade 1 group ( n=24), grade 2 group ( n=19) and grade 3 group ( n=17), and according to the micro analytical rating scale (MARS), the CMBs patients were divided into the lobar group ( n=19), the deep or infratentorial group ( n=17) and the mixed group ( n=24).The relationship between serum MMP-9 level and the location and severity of CMBs was analyzed. SPSS 19.0 software was used for data statistical analysis.One-way ANOVA, t-test and rank sum test were used for comparison. Logistic regression analysis was used to analyze the influencing factors. Pearson correlation analysis and Spearman correlation analysis were used for correlation analysis. Results:The level of MMP-9 in CMBs group was significantly higher than that in control group (208.13(142.25, 285.88) μg/L, 149.50(93.40, 186.51)μg/L), and the difference was statistically significant ( P<0.05). Serum MMP-9 level was a risk factor of CMBs ( β=1.322, OR=3.750, 95% CI=2.038-7.997, P=0.002). The difference of level of MMP-9 in different severity of CMBs was statistically significant (147.55(109.25, 266.47)μg/L, 242.12(147.55, 288.80)μg/L, 270.42(203.43, 364.27)μg/L, P=0.017). Serum MMP-9 level was positively correlated with the number of CMBs ( r=0.371, P=0.003). The difference of MMP-9 level of CMBs in different locations were statistically significant (249.77(158.43, 338.46)μg/L, 188.83(138.52, 243.15)μg/L, 210.65(144.25, 255.78)μg/L, P=0.013). The increased serum MMP-9 level was a risk factor for CMBs( β=0.401, OR=1.122, 95% CI=1.004-1.204, P=0.036). Conclusion:The increased level of serum MMP-9 may be a risk factor of CMBs, especially for CMBs in cerebral lobesand, and the level of MMP-9 is positively correlated with the severity of CMBs.

Objective:To establish the "1+3" management module of high-value consumables in Operation Room and verify its application, so as to provide new ideas for cost management of consumables in Operation Room.Methods:The Operating Room team of Shenzhen Hospital of Southern Medical University designed a "1+3" management module in 2022, where "1" referred to the management process of high-value consumables in Operation Room, and "3" referred to the precise management of consumables in Operation Room warehouse, the management of closed-loop use of Operation Room consumables and adverse event management of consumables. Surgeries using high-value consumables in the Thoracic Surgery Department, Gastrointestinal Surgery Department, and Urology Department of the hospital were selected as the research objects. The surgeries using conventional consumables from January to June 2022 were set as the control group, and the surgeries implementing the "1+3" management module from July to December 2022 were set as the observation group. The number of consumables received by the itinerant nurses before the operation and the number of high-value consumables returned after the operation were compared between the two groups. And the number of missed and error charges for high-value consumables in the two groups were counted and compared.Results:The number of consumables received before operation in the control group was higher than that in the observation group, and the difference was statistically significant ( P<0.05). The number of high-value consumables returned in the observation group was less than that in the control group, and the difference was statistically significant ( P<0.01). The proportion of missed charges for consumables in the observation group was lower than that in the control group, and the difference was statistically significant ( P<0.01), but there was no statistically significant difference in the proportion of incorrect charges between the two groups ( P>0.05) . Conclusions:The "1+3" management module for high-value consumables in Operation Room makes the process of receiving, returning, and charging high-value consumables clear, with traceable data, achieving refined management of high-value consumables in Operation Room, reducing the number of high-value consumables returned to the warehouse and reducing the proportion of missed consumables, which is conducive to effective cost control in Operation Room.

Objective:To investigate the impact of small extracellular vesicles(sEVs) derived from perirenal adipose cells on the biological behavior of renal tubular epithelial cells under diabetic conditions and the underlying molecular mechanisms.Methods:Primary perirenal adipose cells were extracted from db/m and db/db mice for in vitro culture. The culture supernatant was collected and sEVs(NDM-sEVs PRAT-Adipo, DM-sEVs PRAT-Adipo) were extracted by ultracentrifugation. The sEVs were incubated with human renal tubular epithelial cell line(HK-2) to observe changes in their proliferation, apoptosis, autophagy, and epithelial-mesenchymal transition(EMT) levels. The protein composition of sEVs was analyzed using mass spectrometry to explore the molecular mechanisms. Results:CCK8 results showed that the proliferation level of HK-2 cells after DM-sEVs PRAT-Adipo intervention did not change significantly compared with the two control groups(Ctrl group and NDM-sEVs PRAT-Adipo intervention group). Western Blot(WB) results indicated that there were no significant changes in apoptosis levels(Bcl-2, Cleaved-caspase 3, Caspase 3) and autophagy levels(p62, LC3BⅠ, LC3BⅡ) in the DM-sEVs PRAT-Adipo intervention group compared with the two control groups. WB and immunofluorescence results demonstrated that DM-sEVs PRAT-Adipo intervention upregulated the expression levels of mesenchymal cell marker proteins(Vimentin, α-SMA, Snail2) and downregulated the expression level of epithelial cell marker protein ZO-1 in HK-2 cells compared with the two control groups. Mass spectrometry analysis of sEVs revealed that the differential proteins between DM-sEVs PRAT-Adipo and NDM-sEVs PRAT-Adipo were enriched in EMT-related pathways. Among them, the enrichment of thrombospondin(THBS1) in DM-sEVs PRAT-Adipo might be involved in the regulation of EMT in HK-2 cells. Conclusion:Under diabetic conditions, sEVs secreted by PRAT-derived adipocytes promote the upregulation of EMT in renal tubular epithelial cells, a process that may be mediated by the enrichment of THBS1 in sEVs.