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.

Objective:To explore the expression level of serum exosomal miR-223-3p in patients with multiple myeloma (MM) and evaluate its clinical application.Methods:A case-control study was performed. Serum samples were collected from the Department of Hematology, Affiliated Hospital of Nantong University from March 2018 to August 2019, including 68 newly diagnosed untreated MM patients (16 in stage Ⅰ, 22 in stage Ⅱ and 30 in stage Ⅲ), 56 healthy controls and 30 patients with recurrent MM in the same period, and follow-up patients who underwent chemotherapy for 3 months after initial diagnosis. Serum exosomes were purified and identified, RNA was extracted, and RT-qPCR was performed to compare the levels of miR-223-3p in serum exosomes between the groups. The correlation between exosomal miR-223-3p levels and clinical pathological data was analyzed, and the diagnostic efficacy was evaluated by ROC curve.Results:Compared with healthy controls (0.92±0.29), the expression level of serum exosomal miR-223-3p in newly diagnosed MM patients (0.52±0.23) decreased significantly ( U=565, P<0.000 1). According to the international staging system of MM, the expression level of serum exosomal miR-223-3p in patients with stage Ⅰ(0.67±0.26) was significantly higher than that in patients with stage Ⅱ (0.47±0.21) and Ⅲ (0.48±0.19) ( U values were 96.5 and 138 respectively, P were all less than 0.05). In addition, the expression level of serum exosomal miR-223-3p increased after 3 months of chemotherapy (0.69±0.19) compared with that before chemotherapy (0.50±0.22) ( U=228.5, P=0.000 8). Compared with the healthy control group (0.84±0.21), the expression level of serum exosomal miR-223-3p in recurrent MM patients (0.65±0.18) was also decreased ( U=244, P=0.002). At the same time, the expression level of serum exosomal miR-223-3p may be related to the low expression of albumin ( U=411.5, P=0.043 1) and bone damage ( U=309, P=0.001). The ROC curves showed that the AUC of serum exosomal miR-223-3p in the diagnosis of MM was 0.853, which was higher than that of β 2-microglobulin (β 2-MG) (AUC=0.805) and albumin (AUC=0.743). The AUC of the three combined detection was 0.918. Conclusions:Serum exosomal miR-223-3p can be used as a potential marker for auxiliary diagnosis of MM, and its combination with β 2-MG and albumin can improve the diagnostic efficacy of MM.

Objective:To explore a method for detecting recombinant human Annexin A11 (ANXA11) in serum exosomes of pancreatic cancer patients, and then primarily evaluate the clinical value of ANXA11 in pancreatic cancer patients.Methods:A prospective study was conducted and serum specimens from 70 patients diagnosed with PC, 15 patients diagnosed with benign pancreatic mass and 70 patients diagnosed with pancreatitis from the Affiliated Hospital of Nantong University were collected from December 2016 to July 2019. 70 healthy subjects during the same period were selected as control group. The abundance of ANXA11 in serum and exosomes-free serum were detected through parallel reaction monitoring (PRM) basing on high-resolution, high-precision mass spectrometer. Dot immunoblotting created by ourselves for detecting ANXA11 in exosomes and then the methodological evaluation were carried out. Levels of ANXA11 in exosomes in all subjects were statistically analyzed. Moreover, the areas under the curve (AUC) of receiver operating characteristic (ROC) curves were adopted to evaluate the diagnostic efficacy of ANXA11, CA19-9, CEA on PC. The relationship between ANXA11 and clinicopathological parameters as well as prognosis of PC patients was analyzed in the next moment. For analysis, the Mann-Whitney U test was used for comparing between either two groups, and the kruskal-wallis test was used for comparison among four groups. Results:The detection of serum exosome ANXA11 has high sensitivity and repeatability by the method of self-established dot immunoblotting. ANXA11 increased most significantly in the PC group, and the difference was statistically significant ( Hc=58.079, P<0.01) compared with other three groups. ROC curve analysis showed that the diagnostic performance of ANXA11(area under the curve (AUC=0.836) was higher than CEA (AUC=0.656) and equal to CA19-9 (AUC=0.870). The combination of ANXA11 and CA19-9 could improve the sensitivity of diagnosing PC and maintain good specificity. The level of serum exosome ANXA11 before treatment in PC patients was not related to age, gender, tumor size, tumor growth site, lymph node metastasis, distant metastasis and TNM stage ( Z values are 0.052,-0.285,-0.402,0.324,0.888,0.658,1.734, P>0.05). Furthermore, during the 10th day after surgical treatment, the level of ANXA11 showed no statistical difference compared with that before surgery ( Z value is -1.569, P=0.12). The survival time of PC patients was related to the presence of lymph node metastasis, distant metastasis, TNM staging and treatment protocols (χ 2 values are 9.354,6.086,9.389,16.998, P<0.05), while had no correlation with levels of CEA, CA19-9 and ANXA11 (χ 2 values are 1.516, 0.011, 0.159, P>0.05). Conclusions:This study successfully established an original method for detecting ANXA11 levels in serum exosomes of human. Serum exosomes ANXA11 combined with CA19-9 could improve the diagnostic sensitivity of PC.

Objective To evaluate the efficacy of different treatment options for cholecysto-choledocholithiasis (CCL),and try to find the ideal treatment.Methods From January 2006 to January 2016,a total of 3 107 patients with CCL from the Affiliated Hospital of Inner Mongolia Medical University were enrolled.Among them,1 283 patients were in open cholecystectomy (OC) and open common bile duct exploration (OCBDE) group,964 patients were in laparoscopic cholecystectomy (LC) and laparoscopic common bile duct exploration (LCBDE) group,and 860 patients were in endoscopic retrograde cholangiopancreatography (ERCP) + LC group.The clinical data of the three groups were analysed.One-way analysis of variance and chi-square test were performed for statistical analysis.Results From 2006 to 2010,the percentage of patients treated with OC + OCBDE,LC + LCBDE,and ERCP + LC were 56.05％ (829/1 479),25.15％ (372/1 479) and 18.80％ (278/1 479),respectively;from 2011 to 2016,the percentage of patients received the above three treatments were 27.89％ (454/1 628),36.36％ (592/1 628) and 35.75％ (582/1 628),respectively.The difference in the proportion of the same treatment at different times was statistically significant (x2 =4.775,4.168 and 0.669,all P ＜ 0.05).The success rate of surgery in the OC + OCBDE group was 100.00％ (1 370/1 370);while the success rate of surgery in the LC + LCBDE group was 94.26％ (920/976),and 56 patients converted to OC + OCBDE;the success rate of surgery in the ERCP + LC group was 95.00％ (817/860),and 31 patients converted to OC + OCBDE,and 12 patients to LC + LCBDE.The intraoperative complication in OC + OCBDE,LC + LCBDE and ERCP + LC were 2.85％ (39/1 370),3.48％ (32/920) and 1.22％ (10/817),respectively.The incidence rates of postoperative complication were 4.89％ (67/1 370),5.34％ (50/920) and 5.51％ (45/817),respectively.The incidence rates of intraoperative complication of the ERCP + LC group was lower than that of OC + OCBDE group and LC + LCBDE group,and the differences were statistically significant (x2 =6.203 and 3.001;both P ＜ 0.05).However there was no significant difference in incidence rate of postoperative complications among the three groups (all P ＞ 0.05).The hospital stay of the OC + OCBDE group,the LC + LCBDE group and the ERCP + LC group were (6.7 ± 1.3) days,(5.6 ± 1.2) days and (10.9 ± 1.6) days,respectively,and the differences were statistically significant (F =90.010,P ＜ 0.01).The hospitalization expenses of OC + OCBDE group,LC + LCBDE group and ERCP+LC group were (13 720±1 910) yuan,(18 150±1 490) yuan and (25 830 ± 2 430) yuan,respectively,and the differences were statistically significant (F =302.991,P ＜ 0.01).Conclusion The first choice of patients with CCL is endoscopic minimally invasive treatment and open surgery can be used as a remedial method for endoscopic treatment.

Acute radiation exposure usually causes severe dysfunction in various tissues of the organism, among which hematopoiesis, digestion, skin, cardiovascular and nervous system are most obviously affected, often leading to extensive apoptosis and acute radiation syndrome. Apoptosis inhibition or loss could increase proliferation of tumor cells through p53 and NF-κB pathways. Toll-like receptors ( TLRs) are a class of protein molecules involved in non-specific immunity. Up-regulation of the NF-κB pathway results in a significant increase in the radioresistance of cells. TLR4 has a basal cell radiation damage protection effect. Traditional TLR4 agonist lipopolysaccharide is limited in clinical application due to severe toxicity. Recently, some new TLRs agonists have been reported, which have protective effects against the lethal effects of ionizing radiation and lower toxic side effects. Among them, TLR2, TLR4, TLR5 and TLR9 play key roles in radiation protection, such as TLR2 receptor agonist bacterial lipoprotein (BLP), TLR4 receptor agonist heat-activated Salmonella typhimurium (HKST), TLR5 receptor agonism Bacterial flagellin and TLR9 receptor agonist. However, those different TLRs recognize specific components of the pathogen and thus initiate different downstream signaling pathways with different protective effects.

Objective To investigate the clinical diagnostic value of circulating tumor cells (CTCs)and circulating cell-free DNA (cfDNA) in peripheral blood samples in breast cancer. Methods From July 2017 to April 2018, 47 patients with BMC (7 in stage Ⅱ, 19 in stage Ⅲ and 21 in stage Ⅳ), 24 patients with benign breast diseases and 28 healthy people were selected. After collecting peripheral blood samples, serum and blood cells were separated. The size-based high-throughput microfluidic chip was used to capture CTCs. The real-time fluorescent quantitative PCR based on Alu sequence was used to detect the length of cfDNA(247 bp, 115 bp)in the serum, and the ratio of amplified products of long and short fragments was used as the index of DNA integrity. The Mann-Whitney U test or Kruskal-Wallis H test was used to compare the differences between the groups and analyze the relationship between CTCs and cfDNA and clinical parameters of breast cancer. The ROC curve was drawn and the area under the curve (AUC) was used to evaluate the feasibility of blood cell CTCs and plasma cfDNA detection as diagnostic criteria. Results The CTCs and cfDNA of 47 BMC patients were analyzed. The CTCs and cfDNA integrity index (Alu 247/115) of BMC patients were significantly higher than those of physical examination patients[(13.98± 12.36)cells / ml vs (1.14 ± 1.35) cells / ml; 0.7687 ± 0.3868 vs 0.5094 ± 0.2456], and the difference was statistically significant(the U value was 126.5,359.0;P<0.001), the area under ROC curve of CTCs was 0.885 (95％CI: 0.805-0.965), cut-off value was 7.68/ml, sensitivity was 80.4％, specificity was 96.4％. The area under ROC curve of Alu 247/115 was 0.727(95％CI: 0.608-0.847), cut-off value was 0.431, sensitivity was 71.7％, specificity was 71.4％. The AUC of CTCs and Alu 247/115 was 0.919 (95％CI 0.854-0.984), which was higher than the single test of each indicator. Conclusions CTCs and cfDNA may be the potential biological indicators for breast cancer diagnosis. The combined detection of CTCs and cfDNA maybe improve the diagnosis rate of breast cancer patients.

Objective To explore the relationship between adipocyte fatty acid binding protein (A-FABP) and lower limb vascular disease (LLVD) in the elderly with type 2 diabetes(T2DM).Methods Bilateral lower limb vessels were checked by the High Resolution Color Doppler in all the subjects,including 40 healthy subjects as control (group A),126 T2DM patients.42 T2DM patients had no LLVD (group B),40 had mild LLVD (group C),and 44 had severe LLVD (group D).The levels of plasma A-FABP,blood glucose,lipid profiles,HOMA-IR,hypersensitive C reactive protein (hs CRP),and e-glomerular filtration rate were determined.Results The levels of plasma A-FABP were in the following ascending order of group A(4.5± 1.7)μg/L＜group B(6.1±2.1)μg/L＜group C (7.2 ± 2.3)μg/L ＜ group D (8.4 ± 3.2)μg/L (P＜ 0.01).A-FABP levels elevated along with the decrease of ABI and the increase of L IMT.Multiple logistic regression analysis showed that A FABP levels was main influencing factor of lower limb vascular disease (LLVD) in the elderly with type 2 diabetes.LDL C,HbA1c,HOMR-IR,hs-CRP were the predictive factors for the plasma A-FABP levels in the elderly with T2DM after multiple stepwise regression analysis.Conclusions For elder T2DM patients,the level of plasma A FABP is correlated with the degree of LLVD,and plays an important role in the progress of LLVD.

OBJECTIVETo investigate the safety of different level of tween 80 by comparing the degree of pseudoanaphylactoid reactions (PR) induced by medicinal tween 80 and injectable tween 80.

METHODThe analysis of vascular permeability of the mice ears: ICR mouse were divided into different test groups, the mice were intravenously injected with solutions of medicinal tween 80 and injectable tween 80 with 0.2%, 1% and 5% concentration, positive control Compound 48/80 and 5% glucose injection. All test substances were mixed with 0.4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after injection. The analysis of vascular permeability of the rat's skin: the rats were intravenous injected with 0. 6% Evans blue normal saline solution first, 10 minutes later, the same substances were intradermal administrated into the back of rats. The rats were sacrificed and the diameter of locus ceruleus and the content of Evans blue leak out were measured 20 min after injection.

RESULTMedicinal tween 80 and injectable tween 80 with 5% concentration caused obvious vascular hyper permeability in ICR mice, but the degree of vascular hyperpermeability caused by injectable tween 80 was lighter than by medicinal tween 80. Other tween 80 didn't cause obvious vascular hyper permeability in the ears of mouse. The solution of different concentration of tween 80 caused obvious locus ceruleus reaction in rat's back. As for the content Evans blue leak out, there was no statistical significance between each group except positive control Compound 48/80 group.

CONCLUSIONTween 80 can cause obvious vascular hyper permeability and the effect is dose dependent, which indicated that tween 80 can cause PR. On the other hand, injectable tween 80 is more security than medicinal tween 80, the dosage of tween 80 should be still controlled strictly so that to decrease the incidence of PR.

Anaphylaxis ; chemically induced ; Animals ; Injections ; methods ; Male ; Mice ; Mice, Inbred ICR ; Polysorbates ; toxicity ; Rats ; Rats, Wistar

OBJECTIVETo compare the anaphylactoid effect of old Shengmai injection and new Shengmai injection on Cynomolgus monkey.

METHODCynomolgus monkeys were randomly divided into 4 groups, and were respectively injected with 5% glucose injection, old Shengmai injection, new Shengmai injection, positive control drug. The changes of each monkey were observed from injection before until 24 hours after injection, and the response level was determined according to the severity of the symptoms. Blood samples were collected before injection and at 10 min after injection for measuring histamine content in plasma. Blood pressure and heart rates were detected before injection and at 10 min after injection. Sensitization of the injection was comprehensively determined by combined the response level of symptoms and the histamine level.

RESULTThe Cynomolgus monkeys administrated with old Shengmai injection showed typical symptoms of anaphylactoid reactions and the content of serum histamine is not more than doubled. The Cynomolgus monkeys administrated with new Shengmai injection showed untypical symptoms of anaphylactoid reactions and the content of serum histamine did not rise.

CONCLUSIONThe old Shengmai injection can induce typical anaphylactoid reactions on Cynomolgus monkeys, and the sensitization ability is strong. The symptoms of anaphylactoid reactions induced by the new Shengmai injection appeared later and showed lesser degree with the sensitization lower.

Anaphylaxis ; chemically induced ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; toxicity ; Injections ; methods ; Macaca fascicularis ; Male

OBJECTIVETo evaluate the sensitization effect of different components of Shengmai injection (new production process) on Beagle dogs.

METHODBeagle dogs were randomly divided into 7 groups, 3 in each group. Each group was respectively injected with 5% glucose injection, Ginseng Radix et Rhizoma Rubra extract, Ophiopogonis Radix extract, Schisandrae Chinensis Fructus extract, Schisandrae Chinersis Fructus distillate, Shengmaifang, 0.2% tween 80. The changes of each dog were observed from injection before until 24 hours after injection, and the response level was determined according to the severity of the symptoms. Blood samples were collected before injection and at 10 min after injection for measuring histamine content in plasma by ELISA. Sensitization of the injection was comprehensively determined by combined the response level of symptoms and the histamine level.

RESULTOne dog of Ginseng Radix et Rhizoma Rubra extract group showed untypical symptoms of anaphylactoid reactions, and serum histamine of two dogs increased more than doubled. The Beagle dogs administrated with 0.2% tween 80 showed typical symptoms of anaphylactoid reactions, while there was no significant increase of serum histamine. Other groups were observed with no typical anaphylactoid reactions.

CONCLUSIONThe sensitization effect of Shengmai injection (new production process) may be associated with Ginseng Radix et Rhizoma Rubra extract and 0.2% tween 80.

Anaphylaxis ; chemically induced ; Animals ; Dogs ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; toxicity ; Female ; Injections ; Male ; Random Allocation