Objective To investigate the effect of anti-sense osteopontin(ANOPN)on proliferation and metastasis of esophagus cancer cells.Methods An OPN gene recombinant expression vector plasmid was constructed by RT-PCR from human umbilical vein endothelial cell gene and cloned into a mammalian expression vector pcDNA3.1(+).PcDNA3.1-ANOPN was introduced by LipofectinTM.Positive cell clones(ECA-ANOPN),vector-transfected cells ECA-vect and blank cell ECA were used as three groups.RT-PCR and immunocytochemistry assay were used to investigate the expressions of OPN mRNA and protein.The metastasis characteristics of cells were studied by Transwell method.Results The vector was constructed successfully,the sequencing result was identical with that reported in GenBank.Compared with vector-transfected cells(ECA-vect cells) and ECA cells,the growth rate of ECA-ANOPN cells was significantly slowed(P

Objective To analyze the clinical manifestations,treatment and prognosis of adult medulloblastoma (MB).Methods A total of 163 cases of adult MB,confirmed by surgical and pathological diagnosis,were retrospectively analyzed about the clinical manifestations,imaging,treatment and prognosis.There are 108 males and 55 females in the group,whose average age was 28.6-year-old.Results The main clinical manifestations include headache,nausea,vomiting,dizziness,gait disturbance,hypopsia,diplopia,hearing loss and cerebellum crisis.Gross total resection was achieved in 90 cases,subtotal resection in 67 cases and biopsy in 6 cases.Survival time from surgery to progression or death or the last date of follow-up were measured and estimated by the Kaplan-Meier method.Among a total of 160 follow-up patients with MB,postoperative overall survival time was from 7 to 170 months,median survival time was (127±6) months (95％ CI 115.3-138.68).The 5-year overall survival time of adult MB patients was 73.1％ (120/163).The log-rank test was used to compare the significance of the following prognostic variables.Among all clinical factors,patients undergoing the craniospinal irradiation had a significantly better survival rate than those without this treatment (44 months vs 34 months,x2 =8.712,P =0.003).Conclusion Good therapeutic effect of MB be achieved by adopting surgical resection with early enough craniospinal irradiation.Recurrence and metastasis are the two main factors for bad prognosis in adult MB.

Objective To investigate the effect of moderate treadmill exercise together with modified hydroxyapatite chitosan composite hydrogel (CS/HA-g-CS) implantation on repair of full-thickness defects of articular cartilage in rats.Methods Full-thickness cartilage defects were drilled in the patellar groove of bilateral femoral condyles in a total of 24 male SD rats before they were randomly assigned into 4 groups.The control group (BC group) was subjected to no exercise or CS/HA-g-CS implantation;the chitosan group (CHI group) to CS/HA-g-CS implantation without exercise;the moderate treadmill exercise group (MIR group) to exercise 4 weeks after modeling without CS/HA-g-CS implantation;the CHI + MIR group to moderate treadmill exercise plus CS/HA-g-CS implantation 4 weeks after modeling.Half of the animals were sacrificed at week 8 and half at week 16 after operation.Femoral condyles were harvested for gross observation and histochemical measurement by O' Driscoll scoring system.mRNA expressions of glycosaminoglycan,collagen type Ⅱ and BMP-2 were detected by RT-PCR.Results Gross observation revealed:at 8 weeks after modeling,the CHI + MIR group was significantly better than the other 3 groups,with the BC group in the poorest (P ＜ 0.05);at 16 weeks after modeling,the BC group was significantly poorer than the other 3 groups (P ＜0.05) among which there were no significant differences (P ＞ 0.05).O 'Driscoll scoring revealed:at both 8 and 16 weeks after modeling,the CHI + MIR group was significantly better than the other 2 groups and the BC group significantly poorer than the other 3 groups (P ＜ 0.05) but there was no significant difference between the MIR and CHI + MIR groups(P ＞ 0.05).The expressions of collagen type Ⅱ,glycosaminoglycan and BMP-2 were significantly higher in the CHI + MIR group than in the other 3 groups (P ＜ 0.05) and significantly lower in the BC group than in the other 3 groups (P ＜ 0.05).Conclusions Moderate treadmill exercise together with CS/HA-g-CS implantation has significant positive effects on repair of full-thickness defects of articular cartilage in rats than merely moderate treadmill exercise or CS/HA-g-CS implantation alone.The defective cartilage repaired by moderate treadmill exercise together with CS/HA-g-CS implantation contains more collagen type Ⅱ and glycosaminoglycan and shows morphology of nearly normal cartilage.

Objective To make a comparison for the neutrophils prepared either by induction of differentiation of myeloid leuke-mia cell line,or by separation and purification of peripheral blood cells,or by induction of myeloid differentiation of peripheral blood stem cells.Methods NB4 cells were induced differentiation by 1μmol/L all-trans retinoic acid (ATRA)to mature granulo-cytes;neutrophils were separated and purified from peripheral blood by lysis of red blood cells followed by negative selection using magnetic bead-labeled antibodies;hematopoietic stem cells were separated and purified from peripheral blood by Percoll gradient centrifugation followed by negative selection using magnetic bead-labeled antibodies,and were induced to myeloid differentiation by GM-CSF and G-CSF.Morphology and purity of neutrophils prepared by these three methods were studied by means of MGG stai-ning.CD18 protein expression and subcellular distribution were studied by means of immunofluorescence staining.Results Purity of neutrophil was above 40% by induction of differentiation of NB4 cells,and was about 90% if purified from peripheral blood,and was above 70% if induced by myeloid differentiation of peripheral blood stem cells.There was no obvious difference for CD18 ex-pression in neutrophils prepared by these three methods,and staining of CD18 had a dotted pattern distributed in these cells.Con-clusion Peripheral blood neutrophils prepared by lysis of red blood cells followed by negative selection using magnetic bead-labeled antibodies are with high purity and viability which is suitable for immediate test of neutrophils from fresh blood.Neutrophils pre-pared by myeloid differentiation of hematopoietic stem cell are with high viability and last for days,which can be used in long test for neutrphils.

NS2 is a nonstructural protein of Periplaneta fuliginosa densovirus(PfDNV) with a molecular mass of 30 kD,whose function is not yet clearly understood. In order to study the expression,subcellular distribution and the function of NS2 protein,the coding region of NS2 was amplified from the hindgut tissue of cockroaches infected with PfDNV by RT-PCR and then the recombinant prokaryotic expression vector pET28a-NS2 was constructed. The recombinant plasmid was transformed into E. coli BL21(DE3) to express the 6?His fusion protein in the bacteria. After purification,the fusion protein was injected into New Zealand rabbits to prepare polyclonal antibody. The specificity of the anti-NS2 antibody was successfullyproved by western blotting on the eukaryotic expressed products of NS2 protein.Meanwhile,the full sequence of ns2 gene was also cloned into the eukaryotic expression vector pAC. The recombinant plasmid pAC-NS2 was then transfected into Schneider line 2(S2) cells to express NS2 protein in the insect cells. The subcellular localization of NS2 in the insect cells was then investigated by indirect immunofluorescence technique using the anti-NS2 polyclonal antiserum. The confocal laser scanning microscope observation showed that NS2 protein was located primarily in the cytoplasm with some punctate nuclear staining.

Objective:To study the correlation between the quartering of nerve root subsidence sign (NRS) and the cross-sectional area (CSA) of the narrow segment thecal sac in patients with lumbar spinal stenosis (LSS).Methods:The data of 203 LSS patients in the Fourth People′s Hospital of Hengshui from January 2020 to December 2021 were retrospectively analyzed. All patients underwent MRI cross sectional scanning. The patients were divided into positive type a group ( n=62), positive type b group ( n=32), positive type c group ( n=51), and negative group ( n=58) by NRS quartering method. The minimum CSA, median sagittal diameter (PAD), and lateral recess sagittal diameter of each group were compared. The correlation between NRS quartering classification and the minimum CSA and related indicators of lumbar spinal stenosis was analyzed. Results:The minimum CSA, PAD, and sagittal diameter of the lateral recess in the positive a group, positive b group, and positive c group were all smaller than those in the negative group, while the Visual Analogue Scale (VAS) score and Oswestry Disability Index (ODI) were higher than those in the negative group; The minimum CSA, PAD, and sagittal diameter of the lateral recess in the positive b type and positive c type groups were smaller than those in the positive a type group, while the VAS score and ODI index were higher than those in the positive a type group; The minimum CSA, PAD, and sagittal diameter of the lateral recess in the positive c type group were smaller than those in the positive b type group; The VAS score and ODI index were higher than those of the positive b type group; The differences were statistically significant (all P<0.05). 203 patients were divided into 54 normal cases, 58 mild stenosis cases, 49 moderate stenosis cases, and 42 severe stenosis cases based on the minimum CSA. The coincidence rate between negative NRS and minimal CSA diagnosis as normal was 94.44%(51/54), the coincidence rate between positive type a and minimal CSA diagnosis as mild stenosis was 84.48%(49/58), the coincidence rate between positive type b and minimal CSA diagnosis as moderate stenosis was 53.06%(26/49), and the coincidence rate between positive type c and minimal CSA diagnosis as severe stenosis was 90.48%(38/42). Using the kappa consistency test, the kappa value for quantitative diagnosis of minimum CSA stenosis in NRS and LSS patients was 0.743, indicating good consistency. The kappa values for quantitative diagnosis of NRS, sagittal diameter of lateral recess, and PAD stenosis were 0.271 and 0.335, with poor consistency. NRS typing was negatively correlated with CSA and PAD ( r=-0.723, -0.581, all P<0.001), and positively correlated with VAS score and ODI index ( r=0.473, 0.640, all P<0.001). Conclusions:The NRS quartering method has a good consistency in diagnosing the severity of LSS patients and the minimum CSA of stenosis segments, suggesting that the NRS quartering method can better reflect the degree of Spinal stenosis, which can not only be used as an auxiliary indicator for qualitative diagnosis of LSS, but also has a high value in quantitative diagnosis.

Nonalcoholic fatty liver disease(NAFLD)has become the most common liver disease in the world and is an important risk factor for the progression to hepatocellular carcinoma.However,the pathogenesis of NAFLD remains unclear,and there is still a lack of specific treatment measures.Sterol regulatory element-binding proteins(SREBP)are an important nuclear transcription factor,which mainly maintains the balance of lipid metabolism inside the body by activating the genes associated with the synthesis and uptake of cholesterol,fatty acids,and triglycerides,and therefore,SREBP are a target for the treatment of metabolic diseases.This article reviews the latest advances in SREBP in the pathogenesis of NAFLD and the latest evidence of SREBP-targeted therapy for NAFLD.It is worth noting that recent studies have shown that SREBP inhibition can cause liver injury together with autophagy damage.Therefore,excessive inhibition of lipogenesis may exert a counterproductive effect on the treatment of NAFLD.In conclusion,SREBP is a promising therapeutic target for NAFLD;the molecular mechanism of SREBP in lipid metabolism is regulated by many factors,and these factors are being deeply explored and analyzed,which has an important clinical significance for the treatment of NAFLD.

Objective To explore the application and prospects of artificial intelligence(AI)in metabolic associated fatty liver disease(MAFLD)based on bibliometrics.Methods Relevant litera-ture on the application of AI technology in MAFLD was retrieved from the Web of Science Core Collec-tion(WoSCC)database.Bibliometric analysis was conducted using CiteSpace,VOSviewer,R pack-age"bibliometrix",and online bibliometric analysis was platformed to identify hotspots and trends in this field.Results A total of 303 eligible articles were obtained.Since 2017,the number of papers in this field had experienced explosive growth.The United States was leading the research on the appli-cation of AI in MAFLD and was the most frequent participant in international cooperation.San Diego of University of California was the institution with the highest number of publications.Rohit Loomba was the author with the highest number of publications,having published 14 articles.The co-cited keyword clustering labels revealed 10 major clusters:digital image analysis,machine learning,computer-aided diagnosis,fibrosis stage,automated quantitative analysis,metaproteomics,non-invasive diagnosis,ul-trasonography,electronic health records,and knowledge representation.Current research on the appli-cation of AI in MAFLD mainly focused on the diagnosis,differential diagnosis,and staging of MAFLD.Image recognition and analysis,intelligent assisted diagnosis,AI algorithms,and monitoring disease progression will be important research directions for AI in MAFLD.Conclusion Research on the application of AI in MAFLD is experiencing exponential growth.Given enormous potential and clini-cal application prospects of this field,the application of AI in MAFLD-related areas will remain a re-search hotspot in the future.

Objective To explore the application and prospects of artificial intelligence(AI)in metabolic associated fatty liver disease(MAFLD)based on bibliometrics.Methods Relevant litera-ture on the application of AI technology in MAFLD was retrieved from the Web of Science Core Collec-tion(WoSCC)database.Bibliometric analysis was conducted using CiteSpace,VOSviewer,R pack-age"bibliometrix",and online bibliometric analysis was platformed to identify hotspots and trends in this field.Results A total of 303 eligible articles were obtained.Since 2017,the number of papers in this field had experienced explosive growth.The United States was leading the research on the appli-cation of AI in MAFLD and was the most frequent participant in international cooperation.San Diego of University of California was the institution with the highest number of publications.Rohit Loomba was the author with the highest number of publications,having published 14 articles.The co-cited keyword clustering labels revealed 10 major clusters:digital image analysis,machine learning,computer-aided diagnosis,fibrosis stage,automated quantitative analysis,metaproteomics,non-invasive diagnosis,ul-trasonography,electronic health records,and knowledge representation.Current research on the appli-cation of AI in MAFLD mainly focused on the diagnosis,differential diagnosis,and staging of MAFLD.Image recognition and analysis,intelligent assisted diagnosis,AI algorithms,and monitoring disease progression will be important research directions for AI in MAFLD.Conclusion Research on the application of AI in MAFLD is experiencing exponential growth.Given enormous potential and clini-cal application prospects of this field,the application of AI in MAFLD-related areas will remain a re-search hotspot in the future.

Pulmonary arterial hypertension(PAH)is a complex pulmonary vascular disease characterized by pro-gressive elevation of mean pulmonary artery pressure resulted from the pathological feature of pulmonary vascular re-modeling.Without medical intervention,PAH can eventually lead to right heart failure and death of patients.Up to the present,there are few treatment options for PAH are still mainly function through pulmonary vasodilation.Although these treatments can alleviate symptoms,the prognosis remains poor.In recent years,breakthroughs have been made in understanding the pathogenesis of PAH,thus support the development of new treatment strategies tar-geting at dysregulation of signaling pathways in PAH.This review focuses on five critical pathways and the relevant drugs those entered phase Ⅱ clinical trials and discusses their therapeutic potential,so to provide a basis for future research on targeting therapies for PAH patients.