A novel bivalent anti-c-MET/PD-1 bispecific antibody exhibits potent cytotoxicity against c-MET/PD-L1-positive colorectal cancer.

Previously, we generated a novel bispecific antibody (BsAb) simultaneously targeting both c-MET and PD-1 (PDCD1), which can bridge T cells and c-MET positive tumor cells. However, the specific mechanisms and antitumor activities of the BsAb against c-MET/PD-L1 (CD274) positive colorectal cancer (CRC) is not completely understood. In this study, in addition to the tumor intrinsic mechanism investigation with molecular biology assay in vitro, a humanized mouse model was used to evaluate antitumor activity of the BsAb in vivo. The BsAb could inhibit c-MET/PD-L1+ CRC cell migration and show strong antitumor activity against HCT116 tumors in mice, potentially by inducing the degradation of c-MET protein in a dose and time-dependent manner. The BsAb could suppress the phosphorylation of c-MET downstream proteins GRB2-associated-binding protein 1 (Gab1) and focal adhesion kinase (FAK). Considering the tumor extrinsic mechanism, the BsAb may promote phagocytosis of macrophage. Furthermore, the level of plasma exosomal-c-MET/PD-L1 is able to distinguish CRC patients from healthy controls. In summary, the BsAb exhibited potent anti-tumor activities by two distinguished mechanisms: inhibition of c-MET signal transduction and promotion of macrophage-mediated phagocytosis. Our BsAb may provide a novel therapeutic agent for patients with c-MET/PD-L1+ CRC, and the status of exosomal-c-MET/PD-L1 can serve as a biomarker to predict responsiveness to treatment of our BsAb.

[Implantation of a monofocal intraocular lens after femtosecond laser intrastromal keratotomy and contralateral implantation of a trifocal IOL: a case report].

An age-related cataract patient who underwent femtosecond laser intrastromal keratotomy in the right eye for presbyopia correction 8 years ago was subjected to femtosecond laser-assisted phacoemulsification, with implantation of a monofocal intraocular lens (IOL) and a trifocal IOL in the right and left eyes, respectively. The corneal stromal ring was complementary to the monofocal IOL, which recovered the distance and near visual acuity, in the right eye postoperatively. The trifocal IOL provided good intermediate visual acuity for the left eye. The vision of the patient reached an ideal level for all visual distances. The binocular fusion was within the normal range, and the stereoscopic vision was restored. We hope that this case report can act as a reference for the treatment of cataract after presbyopia corrective surgery.

[Study on the expression of p62 protein in patients with lung adenocarcinoma].

To explore the expression of p62 protein in lung adenocarcinoma (LUAD). In this study, a cross-sectional study was adopted. From December 2011 to May 2013, 60 patients with lung adenocarcinoma who were diagnosed and treated in Tongji Hospital of Tongji University, Shanghai were selected for paraffin embedding and tissue chip preparation, and immunohistochemistry (IHC) technology was used to detect the expression of p62 in lung adenocarcinoma patients' cancer tissues and adjacent tissues, and analyze the relationship between p62 expression and the clinicopathological characteristics and survival prognosis of patients with lung adenocarcinoma; at the same time, 6 cases of lung adenocarcinoma were selected by random sampling cancer tissues and adjacent tissues were detected by Western Blot (WB) to detect p62 protein and analyzed by gray value. Preoperative examination specimens of inpatients with lung adenocarcinoma diagnosed from April 2018 to early October 2019, and plasma specimens of healthy subjects were collected, and enzyme linked immunosorbent assay (ELISA) was used to detect lung adenocarcinoma patients and healthy patients. The expression of p62 in the plasma of the subjects was statistically analyzed using SPSS 22.0 software. The results of IHC showed that the positive expression rate of p62 in cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant (t=5.593, P<0.001). Similarly, WB results showed that the expression of p62 protein in cancer tissues was significantly higher than that in adjacent tissues. It is statistically relevant (t=2.238, P=0.049). The expression of p62 was statistically correlated with tumor size, clinicopathological stage and lymph node metastasis in patients with lung adenocarcinoma (all P<0.05). The overall survival of patients with lung adenocarcinoma with high p62 expression was worse than that of patients with low p62 expression (95%CI was 0.238-0.870, P=0.028), suggesting that the high expression of p62 is related to the poor prognosis of patients with lung adenocarcinoma. The level of p62 protein in the plasma of patients with lung adenocarcinoma was significantly higher than that in the healthy control group. The difference was statistically significant (t=8.533, P<0.001). The area under the receiver operating characteristic curve was 0.835 (95%CI was 0.779-0.891, P<0.001), which is significantly higher than CEA, CA125, CA153 and other single traditional indicators, and the combined detection of four indicators has the highest diagnostic efficiency. p62 was strongly expressed in cancer tissues and serum, which is related to the poor prognosis and overall survival rate of LUAD patients.

[Merkel cell carcinoma: a clinicopathological study of 10 cases].

Objective: To investigate the clinicopathological features, differential diagnosis and prognosis of Merkel cell carcinoma (MCC). Methods: The clinical and pathological data of 10 patients with MCC were collected at the 940th Hospital of PLA. The histological characteristics were examined. Immunohistochemical EnVision method was used to detect thyroid transcription factor-1 (TTF1), broad-spectrum cytokeratin (CKpan), CK20, S-100, Ki-67, CD56, chromogranin A, synaptophysin and other markers in the 10 cases. Results: Intradermal MCC of the skin showed a nested, cord-like, cribriform distribution, polygonal cells, uniform size, and lack of cytoplasm. Tumor cell nuclei were large and round, with clear nuclear membranes, fine and scattered chromatin, absence of nucleoli, and mitotic figures of 10 per 50 high power fields. Among them, one patient had sarcoma and squamous cell carcinoma in situ, one patient had squamous cell carcinoma in situ, and one patient had unique cell morphology. Immunohistochemical staining showed that all cancer cells expressed CKpan, synaptophysin and CD56. There were seven cases with perinuclear dot-like positivity of CK20. Six MCCs expressed chromogranin A to varying degrees, while 2 MCCs were weakly positive for p63. The nuclear positive index in the Ki-67 hotspot area was 60%. Conclusion: The histology of MCC varies. Rendering a correct diagnosis of MCC requires adequate sampling, close correlation with clinical history and rational use of immunohistochemical staining. The treatment requires standardized surgery, postoperative radiotherapy and multimodal chemotherapy. Immunotherapy may replace the traditional treatment in the future.

[Investigation of epidemiological characteristics and influencing factors on dust-exposed working age of pneumoconiosis cases in a city].

Objective: To investigate epidemiological characteristics and influencing factors on dust-exposed working years of pneumoconiosis cases in Yantai. Methods: In January 2020, By Cluster sampling way, Using descriptive statistics to analyze dust-exposed working years of the reported 3307 new cases of pneumoconiosis from 2009 to 2019 in Yantai. Analyzing the variation trends by the chi-square trend test. Analyzing single fator by chi-square test. Using multiple classification Logistic Regression analysis to analyze multiple influencing factors. Results: The dust-exposed working age of pneumoconiosis cases decreased by years (P<0.05) . The starting age of dust exposure of cases increased year by year, while positively correlated with the dust-exposed working age (r=-0.217, P<0.05) . The years of dust exposure, starting age of dust exposure, industry, type of economy and scale of enterprise were main influence factors on the dust-exposed working age of pneumoconiosis cases. Conclusion: We should strengthen the management on prevention and control condition of pneumoconiosis in metal smelting and processing industry and privately-owned, small and micro enterprises, and pay attention to workers starting exposed to dust in the age of ≥28 years old.

Effects of sophorolipids on fungal and oomycete pathogens in relation to pH solubility.

AIMS: The objective of this study was to determine the effects of sophorolipids on several fungal and oomycete plant pathogens and the relationship between sophorolipids at different pH and antimicrobial activities. METHODS AND RESULTS: Sophorolipids had different solubility at different pH with a dramatic increase in solubility when pH was 6 or higher. Inhibition of mycelial growth of Phytophthora infestans by sophorolipids was affected by pH values, showing that when the pH value was higher, the inhibition rate was lower. Sophorolipids inhibited spore germination and mycelial growth of several fungal and oomycete pathogens in vitro including Fusarium sp., F. oxysporum, F. concentricum, Pythium ultimum, Pyricularia oryzae, Rhizoctorzia solani, Alternaria kikuchiana, Gaeumannomyces graminis var. tritici and P. infestans and caused morphological changes in hyphae by microscope observation. Sophorolipids reduced ß-1,3-glucanase activity in mycelia of P. infestans. In greenhouse studies, foliar application of sophorolipids at 3 mg ml-1 reduced severity of late blight of potato caused by P. infestans significantly. CONCLUSION: Sophorolipids influenced spore germination and hyphal tip growth of several plant pathogens and pH solubility of sophorolipids had an effect on their efficacy. Application of sophorolipids reduced late blight disease on potato under greenhouse conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings indicated that sophorolipids have the potential to be developed as a convenient and easy-to-use formulation for managing plant diseases.

[Effects of salinomycin on proliferation and apoptosis of oral squamous cell carcinoma].

OBJECTIVE: To investigate the effects of salinomycin on the proliferation and apoptosis of oral squamous carcinoma cells and to further understand the mechanisms of these effects. METHODS: The human oral squamous carcinoma cell line CAL-27 was cultured in different concentrations of salinomycin and cisplatin. After co-culture with 0, 1, 2, 4, 8, 16 and 32 µmol/L salinomycin or 0, 1.25, 2.5, 5, 10, 20, 40 and 80 µmol/L cisplatin for 24 hours and 48 hours, the proliferation of oral squamous carcinoma cells were detected by cell counting kit-8(CCK-8) assay. After being exposed to 0, 2, 4, 8 µmol/L salinomycin and 0, 5, 10, 20 µmol/L cisplatin for 48 hours, the cell cycle of oral squamous carcinoma cells was detected by flow cytometry assay, and Western blot analysis was performed to analyze the expressions of cysteine-containing aspartate-specific proteases-3(Caspase-3), cysteine-containing aspartate-specific proteases-9(Caspase-9), poly ADP-ribose polymerase (PARP), protein kinase B (Akt) and phosphorylated protein kinase B (p-Akt) protein in oral squamous carcinoma cells. RESULTS: Both salinomycin and cisplatin significantly inhibited the proliferation of oral squamous cell carcinoma CAL-27 cells in a time- and dose-dependent manner. However, compared with the first-line chemotherapeutic drug cisplatin, salinomycin showed stronger anti-proliferation activity in oral squamous carcinoma cells than cisp-latin (P < 0.001). After being exposed to 8 µmol/L salinomycin, CAL-27 cells exhibited markedly higher proportion in quiescent/ first gap phases (40.40%±1.99% vs. 64.46%±0.90%, P < 0.05), and had a significantly lower proportion in synthesis phases and second gap / mitosis phases (24.32%±2.30% vs. 18.73%±0.61%, P < 0.05; 35.01%±1.24% vs. 16.54%±1.31%, P < 0.05) compared with the dimethyl sulfoxide control group; moreover cisplatin didn't show cell-cycle specific effect on CAL-27. Western blot proved that salinomycin could up-regulate the expressions of Caspase-3 and Caspase-9 protein in oral squamous cell carcinoma CAL-27 cells (P < 0.05). At the same time, the levels of PARP, Akt and p-Akt protein were down-regulated (P < 0.05). CONCLUSION: Compared with cisplatin, salinomycin has a better inhibitory effect on the proliferation of oral squamous carcinoma cells and blocks the cell cycle process at the quiescent / first gap phase. At the same time, salinomycin could trigger apoptosis of oral squamous carcinoma cells and the mechanism is associated with the Akt/p-Akt signaling pathway.

[Effect of silent information regulator 1 on the LPS induced lncRNA expression of macrophages in mice].

Objective: To investigate the effect of Silent information regulator 1 (Sirt1) on the expression profile of long non-coding RNA (lncRNA) in macrophages upon lipopolysaccharide (LPS) treatment. Methods: Peritoneal macrophages (PM) were isolated from nine wild-type C57BL/6 male mice (wild-type group) and nine myeloid-specific Sirt1 knock-out mice (knock-out group). RNA samples were extracted from macrophages stimulated with 1 µg/ml LPS. Sequencing and the differentially expressed lncRNA were screened after the RNA was quantified. The threshold set for up-and down-regulated genes was a fold change (wild-type group/knock-out group) ≥2 and P≤0.05. Afterwards, gene ontology (GO) and pathway enrichment analysis were conducted and co-expression network map was constructed. Results: Four hundred and forty five lncRNA genes were differentially expressed (185 lncRNA genes were up-regulated and 260 lncRNA genes were down-regulated). Two hundred mRNA genes were differentially expressed (113 mRNA genes were up-regulated and 87 mRNA genes were down-regulated). It was found that the differentially expressed lncRNA genes and the predicted corresponding target genes were mainly distributed in the regions of biological processes of macrophage inflammatory response, macrophage chemotaxis and cell metabolism by GO and pathway enrichment analysis. Conclusion: lncRNA expression profile changes significantly in LPS induced macrophages isolated from Sirt1 knock out mice, which is closely related to the function of macrophages.

Population structure and genetic diversity of Setosphaeria turcica from corn in Heilongjiang province, China.

AIM: The aims of this study were to identify races and mating types of Setosphaeria turcica causing northern corn leaf blight in Heilongjiang province of China and analyse the genetic diversity of S. turcica isolates using SSR markers. METHODS AND RESULTS: Based on gene-for-gene interactions, 13 races of S. turcica (races 0, 1, 2, 3, 12, 13, 23, 123, N, 1N, 12N, 3N and 23N) were isolated from infected corn plants in Heilongjiang province. Races 0 and 1 were the predominant races, and race 23N was identified for the first time in the region. Using two pairs of specific primers, three mating types, 'a', 'Aa' and 'A', were identified, with 'a' being the predominant mating type. SSR markers were used to analyse genetic diversity of 60 S. turcica isolates. Five SSR primers were polymorphic, which resulted in 45 reproducible bands with 2-15 bands for each primer. Cluster analysis separated the isolates into five groups at a similarity coefficient of 0·84. Analysis of molecular variance showed that there was significant correlation between SSR groups and mating type of the isolates. No significant correlation was found between SSR groups and physiological races or geographical location of the isolates. CONCLUSIONS: The work reported that races 0 and 1 were the predominant races, and race 23N was identified for the first time in Heilongjiang province with 'a' being the predominant mating type. There was significant correlation between SSR groups and mating type of S. turcica isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results provide information on population structure and genetic diversity of S. turcica causing Northern corn leaf blight, which will facilitate the development of effective disease management programs.

Does audiovisual distraction reduce dental anxiety in children under local anesthesia? A systematic review and meta-analysis.

OBJECTIVES: To perform a systematic review and meta-analysis on the effects of audiovisual distraction on reducing dental anxiety in children during dental treatment under local anesthesia. METHODS: The authors identified eligible reports published through August 2017 by searching PubMed, EMBASE, and Cochrane Central Register of Controlled Trials. Clinical trials that reported the effects of audiovisual distraction on children's physiological measures, self-reports, and behavior rating scales during dental treatment met the minimum inclusion requirements. The authors extracted data and performed a meta-analysis of appropriate articles. RESULTS: Nine eligible trials were included and qualitatively analyzed; some of these trials were also quantitatively analyzed. Among the physiological measures, heart rate or pulse rate was significantly lower (p = .01) in children subjected to audiovisual distraction during dental treatment under local anesthesia than in those who were not; a significant difference in oxygen saturation was not observed. The majority of the studies using self-reports and behavior rating scales suggested that audiovisual distraction was beneficial in reducing anxiety perception and improving children's cooperation during dental treatment. CONCLUSION: The audiovisual distraction approach effectively reduces dental anxiety among children. Therefore, we suggest the use of audiovisual distraction when children need dental treatment under local anesthesia.

Oral amyloidosis: A strategy to differentiate systemic amyloidosis involving the oral cavity and localized amyloidosis.

Amyloidosis is a rare metabolic disorder caused by the extracellular deposition of fibrillar proteins in important organs. The prognosis of oral amyloidosis associated with systemic involvement is generally poor, but is much more favourable in localized cases. Limited information is available regarding the differential diagnosis between localized and systemic amyloidosis involving the oral mucosa, and also the prognosis. In this study, we summarize the strategy for correctly discriminating between localized and systemic oral amyloidosis, to give the right advice during the diagnostic workup.

[Role of PD-1/PD-L1 in microenvironment of breast cancer].

Programmed cell death protein 1 (PD-1), as a checkpoint of the immune signaling pathway, is a hotspot in the field of immuno-oncology. Its binding with ligand (PD-L1) is an important negative regulatory mechanism. Application of PD-1/PD-L1 inhibitors in treatments of breast cancer is still being explored at present. Fully understanding the mechanism of PD-1/PD-L1 in the microenvironment of breast cancer will help their inhibitors play a full role. This review will start with the discovery of PD-1 and PD-L1, describe their main signaling pathways, introduce the common types and applications of PD-1/PD-L1 inhibitors, especially in breast cancer, summarize the basic research of PD-1 and PD-L1 in breast cancer microenvironment in recent years, and seek new strategies of PD-1/PD-L1 inhibitors in breast cancer treatment.

Validation of the Pain Resilience Scale in Chinese-speaking patients with temporomandibular disorders pain.

To validate the Pain Resilience Scale (PRS) for use in Chinese patients with temporomandibular disorders (TMD) pain. According to international guidelines, the original PRS was first translated and cross-culturally adapted to formulate the Chinese version of PRS (PRS-C). A total of 152 patients with TMD pain were recruited to complete series of questionnaires. Reliability of the PRS-C was investigated using internal consistency and test-retest reliability. Validity of the PRS-C was calculated using cross-cultural validity and convergent validity. Cross-cultural validity was evaluated by examining the confirmatory factor analysis (CFA). And convergent validity was examined through correlating the PRS-C scores with scores of 2 commonly used pain-related measures (the Connor-Davidson Resilience Scale [CD-RISC] and the Tampa Scale for Kinesiophobia for Temporomandibular Disorders [TSK-TMD]). The PRS-C had a high internal consistency (Cronbach's alpha = 0.92) and good test-retest reliability (intra-class correlation coefficient [ICC] = 0.81). The CFA supported a 2-factor model for the PRS-C with acceptable fit to the data. The fit indices were chi-square/DF = 2.21, GFI = 0.91, TLI = 0.97, CFI = 0.98 and RMSEA = 0.08. As regards convergent validity, the PRS-C evidenced moderate-to-good relationships with the CD-RISC and the TSK-TMD. The PRS-C shows good psychometric properties and could be considered as a reliable and valid measure to evaluate pain-related resilience in patients with TMD pain.

Short communication: Effect of on-farm feeding practices on rumen protected lysine products.

Two independent studies were conducted to determine whether mechanical mixing of total mixed ration (TMR) or TMR dry matter alters Lys release from 6 rumen-protected Lys (RPL) products (A, B, C, D, E, and F). In the first study, routine mixing procedures were simulated to determine if inclusion of RPL products in TMR altered in situ release of Lys. Following mixing, Dacron bags containing RPL products were ruminally incubated for 0, 6, 12, or 24 h to determine Lys release. The second study occurred independently of the first, in which Lys release from RPL products was evaluated when incorporated into a TMR that differed in dry matter (DM) content. Bags containing TMR and RPL product mixture were stored at room temperature for 0, 6, 18, and 24 h to simulate RPL product exposure to TMR when mixed and delivered once per day. Concentration of free Lys in both studies was determined using ultra-performance liquid chromatography. Following mechanical mixing, ruminal Lys release was significantly greater for C and tended to increase for F. Mechanical mixing did not alter ruminal Lys release from other RPL products evaluated. Hours of ruminal incubation significantly altered Lys release for all products evaluated, and a significant interaction of mechanical mixing and hours of ruminal incubation was observed for A and C. Exposure to lower TMR DM (40.5 versus 51.8%) significantly increased Lys release from B but did not alter Lys release from the other RPL products evaluated. Moreover, time of exposure to TMR significantly increased Lys release from all RPL products evaluated, and a significant interaction of TMR DM and time of exposure to TMR was observed for B and E. These data suggest mechanical mixing and variation in TMR DM may compromise the rumen protection of RPL products; therefore, on-farm feeding practices may alter efficacy of RPL products in dairy rations.

[Retrospective Analysis of 17 Family Homicide Cases].

OBJECTIVES: To summarize the characteristics of family homicide cases and to provide reference for the analysis and prevention of such cases. METHODS: Seventeen solved family homicide cases in Liyang from 2004 to 2014 were investigated. The original registration information, record of scene investigation, corpse inspection report and case situation were analyzed statistically. RESULTS: The characteristics of the 17 family homicides cases showed that most victims were female and most suspects were male, and spouse infidelity and suspected spouse infidelity have higher proportion in the motives for the killings. Murders by patients with psychosis, camouflage murders and murder-suicides occupied a certain proportion in the family homicide cases. CONCLUSIONS: The family homicide cases are correlated with the family factors such as extramarital sexual intercourse and murder by patients with psychosis. Some suspects suicided after murder. The tools for committing crimes have the features of simplicity, randomness and easy source availability.

Effects of corn-based reduced-starch diets using alternative carbohydrate sources on performance of lactating Holstein cows.

Increases in grain prices have led to renewed interest in feeding reduced-starch diets to lactating dairy cows. An experiment was conducted to determine the effects of altering carbohydrate sources and reducing dietary starch on lactational performance, feeding behavior, and ruminal measures of Holstein dairy cows. Fifteen multiparous cows (6 ruminally cannulated) were blocked and assigned to 1 of 5 squares and used in a replicated 3×3 Latin square design with 21-d periods. Cows were fed 1 of 3 experimental diets: a control diet containing 20% brown midrib corn silage, 20% conventional corn silage, and 10% hay crop silage (CON); a reduced-starch high-forage diet containing 53% brown midrib corn silage and 10% hay crop silage (HFOR); and a reduced-starch diet containing the same forages as CON with partial replacement of corn meal by nonforage fiber sources (HNFFS). The CON diet contained (% of dry matter) 26.0% starch and 34.7% neutral detergent fiber (NDF), whereas the HFOR and HNFFS diets contained 21.4 or 21.3% starch and 38.3 or 38.0% NDF, respectively. Dry matter intake tended to be greater for cows fed the CON diet (28.2 kg/d) compared with those fed the HFOR diet (27.2 kg/d). Dry matter intake for cows fed the HNFFS diet was intermediate (27.7 kg/d). Milk yield was greater for cows fed the CON diet (51.6 kg/d) compared with those fed the HFOR diet (48.4 kg/d), but milk fat content tended to increase for cows fed the HFOR diet (3.98%) compared with those fed the CON diet (3.66%). Consequently, fat-corrected and solids-corrected milk yields were unaffected by dietary treatments. Total chewing, eating, and rumination times were similar across all dietary treatments. Rumination time per kilogram of DM was greatest for the HFOR diet, intermediate for the HNFFS diet, and least for the CON diet, whereas rumination time per kilogram of NDF was greatest for the CON diet and least for the HNFFS diet. Mean ruminal pH, NH3-N (mg/dL), and total volatile fatty acid concentrations (mM) were similar across all dietary treatments. Molar proportion of ruminal acetate (mol/100 mol) was increased for cows fed the HFOR diet compared with cows fed the CON diet. Microbial N yield measured by urinary purine derivatives was unaffected by dietary treatment. Reduced-starch diets containing greater amounts of high quality, highly digestible forage or nonforage fiber sources in place of corn meal resulted in similar fat-corrected or solids-corrected milk yield for high-producing dairy cows in the short term.