Triple-negative breast cancer is therapeutically challenging due to the low expression of tumor markers and 'cold' tumor immunosuppressive microenvironment. Here, we present a dual-targeting peptide-drug conjugate (PDC) for tumor inhibition. Our PDC efficiently and selectively delivers cytotoxic Monomethyl Auristatin E (MMAE) into tumor cells via C-X-C chemokine receptor type 4 (CXCR4) and folate receptor 1 (FOLR1) for synergistic inhibition of growth and metastasis. Our results show that the dual-targeting PDC has potent antitumor activity in cultured human cells and several murine transplanted tumor models without apparent toxicity. The combination of dual-targeting PDC and radiotherapy modulates the tumor immunosuppressive microenvironment by increasing CD8⁺ T cell infiltration and attenuating the proportion of myeloid-derived suppressor and regulatory T cells. Therefore, our dual-targeting PDC represents a promising new strategy for cancer therapy that rebalances the immune system and promotes tumor regression.

The comparison between traditional Chinese medicine Jinzhen oral liquid (JZOL) and Western medicine in treating children with acute bronchitis (AB) showed encouraging outcomes. This trial evaluated the efficacy and safety of the JZOL for improving cough and expectoration in children with AB. 480 children were randomly assigned to take JZOL or ambroxol hydrochloride and clenbuterol hydrochloride oral solution for 7 days. The primary outcome was time-to-cough resolution. The median time-to-cough resolution in both groups was 5.0 days and the antitussive onset median time was only 1 day. This randomized controlled trial showed that JZOL was not inferior to cough suppressant and phlegm resolving western medicine in treating cough and sputum and could comprehensively treat respiratory and systemic discomfort symptoms. Combined with clinical trials, the mechanism of JZOL against AB was uncovered by network target analysis, it was found that the pathways in TRP channels like IL-1β/IL1R/TRPV1/TRPA1, NGF/TrkA/TRPV1/TRPA1, and PGE2/EP/PKA/TRPV1/TRPA1 might play important roles. Animal experiments further confirmed that inflammation and the immune regulatory effect of JZOL in the treatment of AB were of vital importance and TRP channels were the key mechanism of action.

OBJECTIVE： To study the effects of Atractylodes macrocephala ethanol extract （AM） on life span of Caenorhabditis elegans（called N 2 nematode for short ），and to investigate its mechanism based on transcription factor SKN- 1/ nuclear factor E 2 related factor 2（Nrf2）. METHODS ：N2 nematode were divided into blank control group ，positive control group （100 μ mol/L curcumin，similarly hereinafter ），AM low-dose ，medium-dose and high-dose groups （100，200，300 μ g/mL， similarly hereinafter ）. The effects of AM on the life span （by average survival time ）of N 2 nematode under normal condition and oxidant stress condition （40 mmol/L H 2O2）as well as its effects on reproductive capability （by the number of filial generation ）of N2 nematode under normal condition were investigated . 700 μmol/L H2O2 was used to establish neuroblastoma cells N 2a oxidant stress model. Effects of positive control ，low-dose，medium-dose and high-dose of AM on the survival rate of model cells were detected by MTT method. After human embryonic renalepithelial cells 293T were transfected with Nrf 2-ARE plasmid ， the effects of positive control and AM on luciferase activity of Nrf2-ARE were detected by luciferase reporter gene method at low，medium and high dose for 24 h and at medium dose for 12，18 and 24 h. RT-PCR was used to detect the effects ofpositive control ，low-dose，medium-dose and high-dose of AM on the mRNA expression of downstream genes NQO- 1 and HO- 1 of Nrf 2 in N 2a cells as well as mRN A expression of en@hactcm.edu.cn downstream genes GCS- 1，GST-7，GST-10，HSP-60，HSP- 16.2 and SOD- 3 of SKN- 1 in N 2 nematode. RESULTS ：Compared with blank control group ，average survival time of N 2 nematode under normal and oxidant stress condition was significantly prolonged in positive control group and AM groups ；the number of filial generation on the first day （except for AM high-dose group ），the number of filial generation on the second day （except for AM low-dose group ） and the total number of filial generation （except for AM low-dose group ） were increased significantly（P＜0.05 or P＜0.01）. The survival rate of N 2a cells in positive control group ，AM medium-dose and high-dose groups were significantly higher than that of model group （P＜0.05 or P＜0.01）. Compared with blank control group ，Nrf2-ARE luciferase relative activity of 293T cells in positive control group and AM groups as well as Nrf 2-ARE luciferase relative activity of 293T cells in AM medium-dose group after different time of treatment were increased significantly （P＜0.01），in dose-dependent and time-dependent trend. Compared with blank control group ，mRNA relative expression of HO- 1 and NQO- 1（except for positive control group ），GCS-1（except for AM low-dose group ），GST-7（except for positive control group and AM low-dose group ）， GST-10 and HSP- 60（except for AM low-dose group ），HSP-16.2（except for positive control group and AM low-dose group ）and SOD-3 （except for positive control group and AM low-dose group ） were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS：AM can prolong the life span of N 2 nematode under normal and oxidant stress condition and improve the its reproductive capacity ，the mechanism of which may be associated with the activation of SKN- 1/Nrf2 signaling pathway.

Objective:To identify and analyze different proteins expression in the periosteum of congenital pseudarthrosis of the tibia (CPT) using tandem mass tags (TMT) proteomics.Methods:The samples were divided into three groups, namely CPT with neurofibromatosis type 1 (NF1) group (NF1-CPT group), CPT without NF1 group (nonNF1-CPT group) and control group (patients with open tibial fracture). A fold change ≥1.5 or ≤0.66 and P-value <0.05 was regarded as the threshold to screen differentially expressed proteins (DEPs). Subsequently, bioinformatics resources such as online tools DAVID and STRING were used to conduct GO annotation, KEGG pathways enrichment and protein-protein interaction (PPI) network with DEPs. Results:A total of 347 proteins differentially expressed in NF1-CPT group, 212 of which were up-regulated and 135 down-regulated. We identified 467 DEPs in nonNF1-CPT group, including 281 up-regulated and 186 down-regulated. Among of them, NF1-CPT group and nonNF1-CPT group shared 231 DEPs, except for HLA-DRB1 which increased in NF1-CPT group but decreased in nonNF1-CPT group. The remaining 230 DEPs showed the same expression trend in the two positive groups, including 117 up-regulated and 113 down-regulated. In particular, a total of 116 proteins were altered only in NF1-CPT group, including 94 up-regulated and 22 down-regulated. However, there were 236 proteins altered only in nonNF1-CPT group, including 164 up-regulated and 72 down-regulated. The results indicated that the pathogenesis of NF1-CPT was similar as nonNF1-CPT largely with a few differences. Finally, compared with nonNF1-CPT, there were 47 proteins changed 1.5-fold and P-value <0.05 in NF1-CPT group. Conclusion:The proteins expression in the periosteum of CPT is different from that of normal tibia. The expression of periosteal protein is also different between NF1-CPT and nonNF1-CPT. The present study will deepen our understanding of the pathogenesis of CPT in the protein level.

Objective:To explore the effect of recombinant human syntaxin-4(STX4) on lipopolysaccharide(LPS)-induced injury in islet β-cells(INS-1).Methods:Pancreatic islet β-cells(INS-1) were divided into Control (blank control), LPS (LPS treatment), LPS+ NC (transfection of negative control vector, LPS treatment), and LPS+ STX4 (transfection of pcDNA-STX4, LPS treatment) groups. RT-qPCR and Western blot were used to detect STX4 mRNA and protein expression, flow cytometry to detect apoptosis, DCFHDA method to detect reactive oxygen species(ROS) level, xanthine oxidation method to detect superoxide orgotein dismutase(SOD) level, colorimetric method to detect glutathione peroxidase(GSH-Px) level, ammonium molybdate colorimetric method to detect catalase(CAT) level, thiobarbituric acid method to detect malonaldehyde(MDA) level, ELISA method to detect the level of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and insulin secretion levels under glucose conditions secreted by cells, Western blot method to detect Cleared Caspase-3, Bcl-2 Associated X Protein(Bax), p65 protein expression. After treatment with NF-κB signaling pathway activator, STX4 up-regulated islet β-cell INS-1 was given LPS stimulation, and the same method was used to measure apoptosis, ROS, SOD, GSH-Px, CAT, MDA levels and secreted IL-1β, TNF-α, insulin levels and Cleaved Caspase-3, Bax, p65 protein levels.Results:Compared with the Control group, the expression of STX4 mRNA and protein in islet β cells of the LPS group decreased, the apoptosis rate, ROS level, and MDA levels increased, and the levels of SOD, GSH-Px, and CAT decreased, the levels of IL-1β, TNF-α increased, the level of insulin secreted by the cells decreased, and the expression levels of Cleaved Caspase-3, Bax, and p65 also increased. NF-κB pathway activator treatment reversed the effect of up-regulated STX4 on islet β-cell apoptosis, ROS, SOD, GSH-Px, CAT, MDA levels and secreted IL-1β, TNF-α levels, and Cleaved Caspase-3 , Bax and p65 protein levels.Conclusion:Up-regulation of STX4 alleviated LPS-induced islet β cell oxidative damage, apoptosis and inflammatory factor release. The underlying mechanism might be related to the inhibition of activated NF-κB signaling pathway.

Objective: To detect the expression of miR-373 in osteosarcoma cells and explore its effects on cell proliferation, invasion, and migration. Methods: Human osteosarcoma cell line SJSA-1 and human osteoblast hFOB 1.19 cultured in vitro. Human osteosarcoma cell line SJSA-1 were randomly divided into four groups: blank control group (CK group), negative control group (NC group), miR-373 over-expressed plasmid group (miR-373 mimic group) and miR-373 inhibited plasmid group (miR-373 inhibitor group). Cells were transfected with FuGENE^® HD transfection reagent. After 48 hours of transfection, the relative expression of miR-373 was detected by quantitative real-time polymerase chain reaction (qRT-PCR), cell proliferation by cell counting kit-8 (CCK-8) and cell invasion and migration by Transwell. Results: The relative expression of miR-373 in SJSA-1 cells was significantly higher than that in hFOB-1.19 cells (P<0.05); compared with CK group and NC group, the relative expression of miR-373 in the miR-373 mimic group increased significantly (P<0.05), and the relative expression of miR-373 in the miR-373 inhibitor group was significantly lower than that in the control group (P<0.05); optical density (OD) value of miR-373 mimic group at 24 h, 36 h, and 48 h were significantly higher than that of CK group and NC group (P<0.05), and OD value of miR-373 inhibitor group at 24 h, 36 h, and 48 h were significantly lower than that of CK group and NC group (P<0.05); the number of cell migration in the miR-373 mimic group was significantly higher than that in the CK group and NC group (P<0.05), and the number of cell migration in the miR-373 inhibitor group was significantly lower than that in CK group and NC group (P<0.05); the number of cell migration in the miR-373 mimic group was significantly higher than that in the CK group and NC group (P<0.05), and the number of cell migration in the miR-373 inhibitor group was significantly lower than that in CK group and NC group (P<0.05). Conclusions The expression of miR-373 is up-regulated in osteosarcoma. Overexpression of miR-373 can promote the proliferation, invasion, and migration of osteosarcoma cells, while the low expression of miR-373 can inhibit the proliferation, invasion, and migration of osteosarcoma cells, which may be an important target for the diagnosis and treatment of osteosarcoma.

At present medical disputes still happen sometimes though governments at all levels, health departments and hospitals pay more and more attention to correctly guide patient's behavior, regulate doctor's behavior in diagnosis and treatment, purify therapeutic environment, strengthen legislation and protect the legitimate rights and interests of doctors and patients. The causes of medical disputes are great many, and the fundamental reason is that the medical behavior from the beginning to the end is related to the life and health, naturally accompanying medical risk and hiding dispute, therefore what we ought to do is not to absolutely eliminate the risk, but to reduce the risks and disputes. In this report, to explore the causes of medical disputes and risks and look for ways to reduce them, the survey of questionnaires was carried out and practical cases of medical disputes were analyzed in hospitals. Seventy-seven cases of medical disputes from 2013 to 2015 had been completed by arbitration or court decisions, and the final arbitral ideas were as follows: invalid doctor-patient communication, low technology and insufficient management were the main causes of medical disputes; the survey of 483 questionnaires on doctors and nurses in the hospital showed that in addition to the above 3 reasons, there were other deep reasons, namely heavy working intensity, low quality of patients or their family members, and the insufficient management runs through all the links in the way. Therefore, to reduce medical disputes, the following aspects should be commenced: effective communication between doctors and patients; improving doctors' clinical diagnosis and treatment ability; optimizing medical management; correcting medical work attitude;timely medical consultation; attaching importance to medical records; doctor's order leaving some leeway or allowing for unpredictable circumstances; constructing healthy hospital culture; paying attention to the physical and mental health of medical staff; actively improving the medical dispute settlement mechanism and related legal system construction.

Objective To study the clinical features of patients with scrub typhus and provide scientific basis for clinical diagnosis and treatment. Methods Clinical data of patients with scrub typhus in the First Affiliated Hospital of Kunming Medical University from January 2016 to December 2016 were collected. Epidemiological data, clinical manifestations, laboratory findings, image examination results, treatment and outcome were retrospectively analyzed. Results The clinical manifestations included 59 cases (100.0％) with fever, 44 cases (74.6％) with headache, 39 cases (66.1％) with chills, 34 cases (57.6％) with muscle and joint pain, 29 cases (49.2％) with prostration, 49 cases (83.1％) with eschar or ulcer, 42 cases (71.2％) with lymphadenectasis, 23 cases (39.0％) with hepatosplenomegaly. Laboratory test results: 51 cases (86.4％) had normal or elevated white blood cell count, 50 cases of eosinophil reduced (84.7％), 27 cases of blood platelet (PLT) reduced (45.8％), 33 cases of albumin reduced (55.9％), 50 cases of alanine aminotransferase (ALT) increased (84.7％), 48 cases of aspartate aminotransferase (AST) increased (81.4％), and 56.1％ (23/41) of the patients with triiodothyronine (T3), thyroxine (T4), free triiodothyronine (FT3) and free thyroxine (FT4) levels significantly lowered, with predominantly free FT3 reduction (82.6％,19/23); C reactive protein (CRP), procalcitonin (PCT), erythrocyte sedimentation rate (ESR) and ferritin were increased in 93.9％(46/49), 35.4％ (17/48), 64.9％ (24/37), and 83.8％ (31/37) of the patients, and 95.6％(43/45) was accompanied with chest radiographic abnormalities. Tetracycline, doxycycline and azithromycin treatment were all effective. Conclusions The clinical manifestations of patients with scrub typhus, involving multisystem, are diverse and thyroid hormones decrease is observed. Early diagnosis and treatment is the key to improve the prognosis of patients with scrub typhus.

Objective To explore the application and significance of multiple clinical pathway training oriented teaching model in clinical teaching of laboratory diagnostic. Methods Totally 50 medical students enrolled in the Second Military Medical University from September to December in 2015 were divided into experimental group and control group. The course consists of theoretical teaching and experi-mental operation. The pathway group (n=25) were introduced into multiple clinical pathway training oriented teaching method. The theoretical teaching was carried out bysimulation examination application, simulation interpretation and simulation diagnosis and treatment, while the experimental course was carried out by using video teaching combined with actual operation. The control group was taught by traditional teaching method using slide teaching and operation display. The theoretical test including case study and operational skill tests were performed among students in both groups after 10 class hours training . The satisfaction questionnaires were conducted to evaluate the effectiveness and satisfaction of teaching guided by clinical pathway. Differences were compared with independent sample t testing using GraphPad Prism 5.0 statistical software. Results The medical records about professional theoretical test including case study and opera-tional skill test in the pathway group were superior to those in the control group with significant statistical difference (both P<0.05). The records of medical students were (81.84±7.21), (42.00±2.79) in the pathway group and (76.24 ±6.98), (37.00 ±3.71) in the control group. The questionnaire result showed that the pathway group's satisfaction was high, especially with the theoretical knowledge andsceneteaching (higher than 80％). The pathway group believed that multiple clinical pathway training helped to improve learning interest and clinical thinking ability . Conclusions Multiple clinical pathway training oriented teaching model is helpful for the medical students to achieve the basic idea of clinical pathway, improve the profes-sional ability, enhance the interest of learning and the quality of teaching, standardize teaching and promote teaching and learning.

Objective The study is to investigate the role of miR-328 in endothelial mesenchymal transition (EMT)induced by high glucose in human umbilical vein endothelial cells (HUVECs)and its signaling mechanism.Methods HUVECs were cultured in high glucose environment to induce EMT;The recombinant lentiviruses were created by miR-328 and antagomiR- 328 transfection of HUVECs.The experiment was divided into seven groups: normal glucose;mannitol group;high glucose;miR-328;miR-328 virus negative control;high glucose + U0126;miR-328 + U0126.Double immunofluorescent staining was used to determine expression of EMT markers;Changes in miR-328 expression is examined by RT-qPCR;The expressions of type Ⅰ/Ⅲ collagen,p-MEK1/2 and p-ERK1/2 are examined by Western blot.Results 1)HUVECs showed positive staining for CD31 and α-SMA in high glucose group.2)The expression of miR-328 was up-regulated(P<0.05)in HUVECs treated by high glucose or miR-328.Compared with high glucose group or miR-328 group,miR-328 expression was less pronounced aftertreatment with U0126.3)The expressions of type Ⅰ/Ⅲ collagen increased in HUVECs treated by high glucose or miR-328(P<0.05) Compared with high glucose group or miR-328 group,typeⅠ/Ⅲ collagen expressions were less pronounced after treatment with U0126.4)The expressions of p-MEK1/2 and p-ERK1/2 were increased in HUVECs treated by high glucose or miR-328 in comparison to the control group (P<0.05);a lower expression of p-MEK1/2 and p-ERK1/2 were observed in U0126 group than in high glucose group or miR-328 group.Conclusions The phenomenon of EMT in HUVECs is induced by high glucose with increased expression of miR-328;overexpression of miR-328 induced EMT in HUVECs;miR-328 induced EMT is related with MEK1/2-ERK1/2 signaling pathway.