Objective To investigate the expressions of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor-C (VEGF-C) and the correlations with clinic parameters,and to make sure the effects of the two genes in the lymphangiogenesis and lymph node metastasis of breast cancer.Methods The expressions of HIF-1α and VEGF-C were investigated by means of immunohistochemistry in samples from 78 cases with breast cancer and 20 cases with breast benign lesions.The density of the lymphatic microvessels immunohistochemically stained by D2-40 antibody was calculated.The relationships between HIF-1α,VEGF-C,LVD and clinicopathological parameters were analyzed statistically.Results HIF-1α expression occurred in 52 out of the 78 tumor samples (66.67％),while VEGF-C expression was observed in 41 out of the 78 tumor samples (52.56％).HIF-1α and VEGF-C expressions in breast cancer were significantly higher than those in beingn disease (15％ and 10％ ; x2 =17.26,P =0.000; x2 =11.71,P =0.001).The expressions of HIF-1α and VEGF-C were significantly correlated with regional lymph nodal involvement (x2 =5.80,P =0.016; x2 =7.26,P =0.007) as well as late TNM classification (x2 =8.51,P=0.004; x2 =6.02,P =0.014),disregarding the patient's age,tumor diameter,histological grade and pathologic type (P ＞ 0.05).In addition,HIF-1α expression was positively correlated with VEGF-C expression (r =0.254,P =0.025).Higher LVD was found in both high HIF-1α and high VEGF-C expression cases (t =2.19,P=0.017; t =3.25,P=0.001).Conclusion HIF-1α may play a crucial role in the lymphangiogenesis and lymph node metastasis by regulating the expression of VEGF-C in breast cancer.Therefore,HIF-1α may become a particularly promising target for controlling lymph node metastasis in breast cancer.

To construct “developmental assessment” in the course of emergency nursing and inten-sive care nursing by applying the combination of formative assessment and summative evaluation, the com-bination of qualitative assessment and quantitative assessment and the incorporation of self-assessment and other evaluation. Specifically, this project contains investigation activity (accounting for 10%), operating activity (10%), checking on work attendance and homework evaluation (15%), reflection activity (15%), comprehensive design experiment evaluation (20%) and theoretical evaluation (30%). Process evaluation and self-evaluation are highlighted, especially feedback and improvement after evaluation. Through multi stage and multi form evaluation, the goal of emergency and critical care professional education has been achieved, which has promoted the development of student nurses' quality and overall development.

Objective:To clone swine leukocyte antigen,class II,DO alpha (SLA-DOA) gene from Banna mini-pig inbred line (BMI) and detect its mRNA expression level in 19 important tissues.Methods:The complete eDNA sequence of SLA-DOA gene was cloned by RT-PCR method from BMI and the mRNA expression pattern in BMI important tissues was examined by semi-quantative RT-PCR method.Nucleotide and protein sequences of SIA-DOA were used to carry out bioinformatics analysis and construct the phylogenetic tree.Results:The eDNA length of BMI SLA-DOA was 1 079 bp,which encoded a protein of 250 amino acids with molecular weight (Mw) 27.81 kD,and isoelectric point (pI) 6.48.Genome structure analysis showed it localized to Sus scrofa chromosomes 7 and consisted of four exons and three introns.Semi-quantitative expression analysis showed that SLA-DOA gene expressed highly in the lymph nodes and stomach;weakly in the heart,skin and duodenum and none in other 14 tissues.Functional bioinformatics analysis indicated that SLA-DOA protein contained one signal poptide,one transmembrane region,three conserved domains,four O-GlcNAc glycosylation sites,18 potential phosphorylation sites and to be located in the cytoplasm with 94.1％ certainty.Phylogenetic analysis demonstrated that BMI (pig) had the closest relationship with cattle.Conclusion:This study have successfully cloned the SLA-DOA gene of Banna mini-pig inbred line,performed bioinformatics analysis and tissue expression profile analysis.It will provide a basis for the studies of BMI xenotransplantation.

Objective To investigate protection by immunization with recombinant Ferritin vaccine of Echinococcus granulosus against protoscolices.Methods ICR mice were randomized into 3groups of 12 mice in each.The mice in group A and B were immunized three times with an interval of two weeks and those in group C did nothing.The animals in all the 3 groups were challenged with 1100 protoscolices intraperitoneally on the 8th week.Serum samples were collected before each inoculation and challenge injection.Seven months later, all the mice were killed and examinated for hydatid cysts.Result The number of cysts was significantly lower in the group A than in group B and C (P＜0.05).The levels of protection afforded were found to be 73% and 85%, respectively.Meanwhile,the number of cysts was markedly lower in group B than in group C(P＜0.05).The rate of protection afforded was 42%.Conclusion Recombinant Ferritin vaccine of Echinococcus granulosus shows partial immune protection.Therefore, it might be a suitable candidate for cocktail vaccine study in the future.

Objective Pathologic complete response (pCR) has been suggested as a surrogate prognostic indicator in breast cancer patients treated with neoadjuvant chemotherapy.We assessed whether the likelihood of pCR and survival is associated with the immunohistochemistry-based molecular subtypes.Methods We retrospectively analyzed the records of l01 patients with breast cancer who received neoadjuvant chemotherapy between January 2007 and January 2010.Patients were dassified into four molecular subtypes based on the immunohistochemistry prnfiles of estrogen receptor,progesterone receptor,and HER2.Logistic regression was used to analyze variables associated with pCR.Results The pCR was achieved in 19 patients (18.8％).The triple negative subtype was an independent predictive factor for pCR (odds ratio,3.35,95％ confidence interval,1.25-9.79,P =0.012),and the Her-2 subtype showed a trend for higher pCR rates (odds ratio,3.11;95％ confidence interval,1.09-10.89,P =0.021) compared with the luminal A subtype.The pCR was significantly associated with prolonged disease-free survival (P =0.002).The triple negative subgroup had shorter disease-free survival (P =0.0006) and overall survival (P =0.008) than the other subgroups.Conclusions We demonstrated that the triple negative and Her-2 subtypes are more likely to obtain pCR when neoadjuvant chemotherapy is given,compared to the luminal A subtype.Despite the high pCR rate,the triple negative subtype showed worse survival outcomes,paradoxically,primarily due to patients who had residual disease.

Objective To establish a method of identification of DKO mouse model of Duchenne muscular dystro-phy, and to assess the dystrophin regeneration after stem cell transplantation.Methods Heterozygous mice were mated and the resulting offspring were used to identify their genotype by SSP-PCR.The plasma creatine kinase level was measured by biochemical analyzer and histological changes in the DKO mice were analyzed using HE staining.Human umbilical cord mesenchymal stem cells were prepared and injected into the DKO mice hindlimb muscle, and dystrophin expression was de-tected by immunofluorescence staining at 2 months after injection.Results Mating of heterozygous mice generated three kinds of genotype offsprings, and 21.2%of the offsprings were identified as DKO genotype (285 bp) .DKO mice showed dystrophic symptoms, their plasma creatine kinase level was as high as 16988.52 ±617.48 IU/L, and significant histologi-cal changes including diverse myocyte sizes, numerous centrally nucleated cells and connective tissue proliferation or in-flammatory cells infiltration.Human dystrophin expression was detected in the DKO mouse hindlimb muscle at two months after injection of human umbilical cord mesenchymal stem cells.Conclusion DKO mouse genotype can be identified by SSP-PCR, and DKO mouse is an ideal animal model for studies of stem cell therapy for Duchenne muscular dystrophy.

Objective:To explore the current situation and influencing factors of forgiveness tendency of clinical nurses.Methods:This study was a cross-sectional survey. From September 2020 to March 2021, a total of 236 clinical nurses from 3 ClassⅢ Grade A hospitals in Yantai were selected by the convenient sampling method. General information questionnaire, the Heartland Forgiveness Scale (HFS) , the Chinese version of the Jefferson Scale of Physician Empathy (JSPE-HP) and Subjective Well-Being (SWB) were used for investigation. Single factor analysis and multiple linear regression analysis were used to analyze the influencing factors of forgiveness tendency of clinical nurses. At the same time, Pearson correlation analysis was used to explore the relationship between forgiveness tendency, empathy and overall well-being. In this study, a total of 236 questionnaires were issued and 227 valid questionnaires were returned. The effective response rate was 96.19%.Results:The total score of HFS of 227 clinical nurses was (113.21±6.94) . The results of multiple linear regression analysis showed that age, educational background, religious belief, empathy level and overall well-being were the main influencing factors of forgiveness tendency of clinical nurses ( P<0.05) . Conclusions:The level of forgiveness tendency of clinical nurses in this study is above medium level. It is necessary to focus on the clinical nurses with low nursing age, low education, low empathy and low happiness and carry out targeted intervention to improve the level of forgiveness tendency of clinical nurses, so as to improve the physical and mental health and nursing quality of clinical nurses.

Artificial joint replacement is an important means for the treatment of severe joint end-stage diseases such as hip and knee joint,which has obtained satisfactory clinical efficacy,but the postoperative periprosthetic osteolysis (PPO),which is mediated by wear particles,restricts the long-term effect of artificial joints.It is found that wear particles increase the expression of cytokines and inflammatory factors by stimulating the cells around the prosthesis,activate different signaling pathways,promote the imbalance between bone formation mediated by local osteoblasts and bone resorption mediated by osteoclast so as to lose of the local bone mass,and eventually produce osteolysis and aseptic loosening.This article reviews the different signal pathways activated by wear particles in recent years,in order to explore the pathogenesis of PPO and to open up new avenues for its prevention and treatment.

Objective:To evaluate the accuracy of the domestic "Skywalker" surgical robot in implementing personalized lower limb alignment reconstruction scheme in total knee arthroplasty (TKA) and the short-term clinical outcome of robotic assisted TKA.Methods:From September 2020 to January 2021, the data of patients who received surgical robot assisted TKA in 5 clinical centers in China (Shanghai Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Southwest Hospital affiliated to Third Military Medical University, The Affiliated Hospital of Qingdao University, The First Affiliated Hospital of Chongqing Medical University and Yantai Yuhuangding Hospital, and Shanghai Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine was the group leader) were prospectively collected and retrospectively analyzed. There were 24 males and 82 females with an average age of 67.6±7.3 years, (range 45-80 years); Average body mass index 26.42±4.31 kg/m 2, all the operation were performed by "Skywalker" surgical robot system according to preoperative design based on CT. The operation time, intraoperative blood loss, hospitalization days and postoperative complications were recorded, and the imaging indexes including hip-knee-ankle (HKA), lateral distal angle of femur (LDFA) and medial proximal angle of tibia (MPTA) measured before and after the operation, implant model indexes (preoperative planning implant model and postoperative implant model) and short-term clinical efficacy indexes [Western Ontario and McMaster Universities (WOMAC) osteoarthritis index] pain score, stiffness score, joint function score, total score and SF-12 score before and 3 months after the operation) were compared. Results:The average follow-up period was 109.60±9.80 d, (range 95-143 d). The average operation time of 106 patients was 105.30±23.22 min; The average intraoperative blood loss was 141.70±58.33 ml; The average length of hospitalization was 5.82±2.80 d. One patient had ischemic stroke after operation, and one patient had abnormal liver function after operation. According to the judgment of the investigator, all of them were not related to the operation. The actual angle error is the difference between the preoperative planning angle and the postoperative measurement angle. The absolute error of 99.1% (105/106) of the HKA angle was within 3°, 90.8% (69/76) of LDFA, 98.7% (75/76) of the MPTA. In 45 patients in one center where data were available, the actual implant models used in all patients were consistent with the preoperative planning size, and there were only differences in version selection such as Asian condyle. WOMAC pain score, joint function score, total score was improved from 7.34±2.85, 25.10±9.85, 34.75±13.02 to 3.34±2.66, 14.68±9.64, 18.66±13.49 before and after operation, respectively, which were statistically significant ( P<0.001) and SF-12 physiological score and psychological score were improved form 27.24±6.42, 30.68±8.26 to 38.83±5.74, 39.36±7.85 before and after operation, respectively, which were statistically significant ( t=7.33, P<0.001; t=4.53, P=0.043). Conclusion:Domestic surgical robot system "Skywalker" can assist the surgeon to achieve accurate and personalized reconstruction of lower limb alignment and achieve satisfactory short-term clinical outcomes. The long-term clinical outcomes of personalized reconstruction and survival rate of implant still need to be further studied.

Objective To investigate the role and related mechanism of the soluble guanylate cyclase(sGC)-cGMP-protein kinase G(PKG)signaling pathway in the amelioration of isoproterenol(ISO)-induced cardiac hypertrophy in mice by aqueous extract of Curcuma kwangsiensis root tubers(GYJS).Methods 72 KM male mice were divided randomly into 6 groups:normal,model,propranolol control(40 mg/kg),and GYJS low-(1 g/kg),medium-(2 g/kg),and high-dose(4 g/kg)groups.Mice in the experimental groups were injected subcutaneously with ISO 10 mg/kg on days 1～3 and ISO 5 mg/kg on days 4～14 to establish a mouse cardiac hypertrophy model.4h after each subcutaneous injection of ISO,the mice in each group were administered the corresponding drugs orally for a dosing cycle of 14 days.The hearts were then removed and whole heart and left ventricle weight were measured.Myocardial tissue pathology was observed using hematoxylin and eosin and Masson staining,and sGC subunit beta-1(GUCY1B3)and transforming growth factor(TGF-β1)were detected by immunohistochemistry.Serum lactate dehydrogenase(LDH),creatine kinase(CK),and Nitric Oxide(NO),and myocardial superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were measured using respective kits.Serum cGMP was detected by enzyme-linked immunosorbent assay and quantitative reverse transcription PCR(RT-qPCR),and atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),GUCY1B3,PKG Ⅰ,and phosphodiesterase(PDE)5A mRNA expression levels were also determined by RT-qPCR.Results Compared with the model group,whole heart and left ventricle weights were significantly reduced in mice treated with propranolol or GYJS(P＜0.001 or P＜0.0001)and myocardial hypertrophy and myocardial fibrosis were significantly reversed.All the treatments significantly elevated myocardial expression of GUCY1B3(P＜0.05 or P＜0.01)and significantly reduced expression of TGF-β1(P＜0.05 or P＜0.01).The myocardial damage markers LDH and CK were significantly reduced(P＜0.05 or P＜0.01)while NO and cGMP were significantly elevated(P＜0.05 or P＜0.01),the myocardial oxidative stress indicator MDA was significantly reduced(P＜0.05 or P＜0.01)and SOD activity was significantly increased(P＜0.05 or P＜0.01).mRNA levels of the myocardial hypertrophy markers ANP,BNP,and PDE5A were significantly reduced(P＜0.05,P＜0.01,or P＜0.001)and the mRNA levels of GUCY1B3 and PKG Ⅰ were significantly increased(P＜0.01 or P＜0.001).Conclusions GYJS may improve cardiac hypertrophy by modulating the sGC-cGMP-PKG signaling pathway.